Compositions for preserving anaerobic microorganisms and methods for making and using same - Patent Application 20070122997

Abstract

Compositions for preserving one or more species of anaerobic microorganisms are provided herein. Also provided are preservation compositions, such as cryopreservation compositions, of anaerobic microorganisms, and methods for preparing and using the preservation compositions.

Description

[Technical field]

[0001] cross reference This application claims the benefit of and priority to U.S. Provisional Application No. 63 / 182,578, filed April 30, 2021, the contents of which are incorporated by reference in their entirety herein.

[0002] The present disclosure relates generally to methods and compositions for preserving anaerobic microorganisms. [Background technology]

[0003] Live biological therapeutic products (LBPs), which contain viable microorganisms, are a class of therapeutic agents being developed for the treatment of a wide variety of medical disorders, including gastrointestinal diseases such as Crohn's disease and ulcerative colitis. LBPs are believed to exert their therapeutic effect through modulation of the gastrointestinal environment, including the microbiome.

[0004] A common approach to deliver live microorganisms to the gastrointestinal tract is via oral delivery of LBPs. However, significant challenges must be addressed when developing LBPs to ensure that they are safe and effective. These challenges include, but are not limited to, maintaining the stability and viability of the microorganisms during manufacture, storage, and administration to achieve the desired therapeutic effect, achieving the desired release profile of the live microorganisms into the gastrointestinal tract, and ensuring that the microorganisms are restored and propagated during passage through the appropriate regions of the gastrointestinal tract.

[0005] Cryopreservation is a technique often used to formulate LBP for oral delivery. Considering that live microorganisms, including anaerobic organisms, may have a short finite life span, the cryopreservation process may facilitate the long period during which the microorganisms remain viable, which is necessary for LBP. However, the use of cryopreservation itself may result in a significant decrease in the viability of live microorganisms, often caused by cell death associated with the formation of intracellular and / or extracellular ice crystals. In addition, the stability of the cryopreservation formulation may also affect the viability during storage, rehydration, and timely recovery of the organism after ingestion by the subject. Although several approaches have been developed to protect microorganisms from damage during the cryopreservation process, including the use of cryoprotectants and lyoprotectants, as well as for the control of the freeze-drying process, to date, these approaches have resulted in LBS with substantially reduced viability, especially over long periods of time, which requires the administration of higher doses of the organism to achieve the desired therapeutic effect.

[0006] Despite efforts made to date, there remains an unmet need to develop preservative compositions, e.g., cryopreservative compositions, that improve the viability of microorganisms during manufacture, storage, and administration, and that facilitate the timely recovery of the organisms after administration, so as to provide a therapeutic effect as the organism traverses the digestive tract of a subject. Summary of the Invention

[0007] The present invention is based, in part, on the discovery of preservation compositions and techniques that promote long-term viability of anaerobic microorgansims and rapid recovery of the microorgansims when ingested by a subject so that the organisms can replicate as they traverse the digestive tract of the subject. As generally described herein, the present disclosure provides compositions for preserving anaerobic microorgansims, including compositions for cryopreserving anaerobic microorgansims. The compositions generally include a viscosity modifier, an energy source for the microorgansims to promote rapid recovery when administered to a subject, and an oxidation-reduction potential, e.g., about -50 mV to about -500 mV, to support the viability of the anaerobic organisms. Additionally, the compositions disclosed herein generally include one or more different types of anaerobic microorganisms, a viscosity modifier, an energy source for the microorgansims, and an oxidation-reduction potential, e.g., about -50 mV to about -500 mV, to support the viability of the anaerobic organisms.

[0008] In one aspect, the disclosure provides a composition (e.g., a cryopreservation composition) for preserving one or more species of anaerobic microorganisms, the composition comprising a viscosity modifier and an energy source for the microorganisms, and when solvated in an aqueous solvent, has a viscosity in the range of about 5 cP to about 6,000 cP, an osmolality in the range of about 300 Osm / L to about 4,000 Osm / L, a pH in the range of about 4 to about 8, and optionally a redox potential in the range of -50 mV to -500 mV.

[0009] In another aspect, the present disclosure provides a cryopreservation composition for cryopreserving one or more species of anaerobic microorganisms, the composition comprising a viscosity modifier and an energy source for the microorganisms, and when solvated in an aqueous solvent, has a viscosity in the range of about 5 cP to about 6,000 cP, an osmolality in the range of about 300 Osm / L to about 4,000 Osm / L, a pH in the range of about 4 to about 8, and optionally a redox potential in the range of -50 mV to -500 mV.

[0010] In each of the foregoing embodiments, the composition has a viscosity in the range of about 50 cP to about 4,000 cP, about 100 cP to about 2,000 cP, or about 100 cP to about 1,000 cP, and / or an osmolality in the range of about 300 Osm / L to about 3,000 Osm / L, about 400 Osm / L to about 3,000 Osm / L, or about 500 Osm / L to about 3,000 Osm / L, and / or a pH in the range of about 4 to about 8, about 5 to about 8, about 4 to about 7.5, about 5 to about 7.5, or about 5.5 to about 7.5.

[0011] In each of the foregoing aspects, the composition of the composition, when solvated in an aqueous solvent, can contain about 0.01% (w / w) to about 0.4% (w / w) of a viscosity modifier. The viscosity modifier can be a polymer composition, such as a composition comprising alginate, acacia, alginic acid, agar, aramic acid, bentonite, carbomer, carrageenan, cellulose, dextrin, gelatin, gellan gum, guar gum, carboxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, methylcellulose, mucin, pectin, starch, xanthan gum, or combinations thereof. In certain embodiments, the polymer composition comprises an alginate, such as sodium alginate, having a viscosity range of, for example, about 250 cP to about 14,000 cP.

[0012] Further, in each of the foregoing compositions, the energy source can be a cell culture medium that allows for the growth of the microorganism. In certain embodiments, the composition comprises about 3% (w / w) to about 15% (w / w) of the energy source when solvated in an aqueous solvent. The energy source can be a monosaccharide, a disaccharide, an oligosaccharide, or a combination thereof. The monosaccharide can be, for example, selected from the group consisting of glucose, fructose, xylose, and combinations thereof. The disaccharide can be, for example, trehalose, sucrose, or a combination thereof. The oligosaccharide can be, for example, an inulin derivative, such as a fructooligosaccharide.

[0013] Further, in each of the foregoing compositions, the composition can further comprise a protein stabilizing agent. In certain embodiments, the composition comprises about 5% (w / w) to about 10% (w / w) of the protein stabilizing agent when solvated in an aqueous solvent. The protein stabilizing agent can be selected from the group consisting of amino acids, antioxidants, disaccharides, reducing agents, polyethylene glycols, and combinations thereof. In certain embodiments, the disaccharide is selected from the group consisting of trehalose, sucrose, isomaltose, lactose, maltose, lactulose, cellobiose, and combinations thereof.

[0014] In certain embodiments, the one or more anaerobic microorganisms (organisms can belong to different genera or species) are present in the form of a cell paste or cell slurry. Depending on the circumstances, in each of the aforementioned compositions, the composition can comprise about 30% (w / w) to about 50% (w / w) of the cell paste or cell slurry when solvated in an aqueous solvent.

[0015] In another aspect, the present disclosure provides a cryopreservation composition for cryopreserving anaerobic microorganisms, the composition comprising (i) about 30% (w / w) to about 50% (w / w) of a cell paste or cell slurry containing one or more anaerobic microorganisms, (ii) about 3% (w / w) to about 15% (w / w) of an energy source for the microorganisms, (iii) about 0.01% (w / w) to about 0.4% (w / w) of a viscosity modifier, and (iv) about 30% (w / w) to about 60% (w / w) of an aqueous solvent, the cryopreservation composition having a viscosity in the range of about 5 cP to about 6,000 cP, an osmolality in the range of about 300 Osm / L to about 4,000 Osm / L, and a pH in the range of about 4 to about 8.

[0016] In another aspect, the present disclosure provides a cryopreservation composition for cryopreserving anaerobic microorganisms, the composition comprising: (i) about 30% (w / w) to about 50% (w / w) cell paste containing one or more anaerobic microorganisms, (ii) about 0.5% (w / w) to about 5% (w / w) raftilose, (iii) about 3% (w / w) to about 10% (w / w) maltodextrin, (iv) about 0.01% (w / w) to about 0.4% (w / w) sodium alginate, and (v) about 5% (w / w) to about 10% (w / w) trehalose, (vi) 0% (w / w) to about 5% (w / w) sucrose, and (vii) about 35% (w / w) to about 55% (w / w) water, the cryopreservation composition having a viscosity in the range of about 5 cP to about 6,000 cP, an osmolality in the range of about 300 Osm / L to about 4,000 Osm / L, and a pH in the range of about 4 to about 8. The composition can have a redox potential in the range of -50 mV to -500 mV.

[0017] In each of the foregoing compositions, the composition can also include a buffering agent.

[0018] The present disclosure further provides a preserved cell composition (e.g., a cryopreserved composition) comprising one or more anaerobic microorganisms, a viscosity modifier, an energy source for the microorganisms, a redox potential in the range of -50 mV to -500 mV, and a water content of less than 5% (w / w). In certain embodiments, the water content is in the range of about 2% (w / w) to about 4% (w / w) or about 2% (w / w) to about 3% (w / w). In certain embodiments, the viscosity modifier is a polymer composition, such as alginate, acacia, alginic acid, agar, aramic acid, bentonite, carbomer, carrageenan, cellulose, dextrin, gelatin, gellan gum, guar gum, carboxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, methyl cellulose, mucin, pectin, starch, xanthan gum, or a combination thereof. In certain embodiments, the polymer composition comprises an alginate, e.g., sodium alginate, e.g., sodium alginate having a viscosity range of 250-14,000 cP. In certain embodiments, the energy source is a cell culture medium that allows for the growth of the microorganism. The energy source can be a monosaccharide, a disaccharide, an oligosaccharide, or a combination thereof. The monosaccharide can be, e.g., selected from the group consisting of glucose, fructose, xylose, and combinations thereof. The disaccharide can be, e.g., selected from the group consisting of trehalose, isomaltose, lactose, maltose, lactulose, cellobiose, sucrose, and combinations thereof. The oligosaccharide can be, e.g., an inulin derivative, e.g., a fructooligosaccharide.

[0019] In certain embodiments, the composition comprises a protein stabilizing agent. The protein stabilizing agent can be selected from the group consisting of amino acids, antioxidants, disaccharides, reducing agents, polyethylene glycols, and combinations thereof. The disaccharides can be selected from the group consisting of, for example, trehalose, sucrose, isomaltose, lactose, maltose, lactulose, cellobiose, and combinations thereof.

[0020] In certain embodiments, a buffer is included.

[0021] In another aspect, the present disclosure provides a cryopreservation composition comprising: (i) one or more anaerobic microorganisms; (ii) an energy source, including, for example, raftilose and maltodextrin; (iii) sodium alginate; (iv) a protein stabilizing agent, including, for example, trehalose, sucrose, and combinations thereof; and (vi) a water content of less than 5% (w / w). The composition may further comprise a redox potential in the range of -50 mV to -500 mV.

[0022] In each of the foregoing compositions, the one or more anaerobic microorganisms may be selected from the group consisting of Akkermansiaceae, Bacillaceae, Bacteroidaceae, Bifidobacteriaceae, Christensenellaceae, Clostridiaceae, Coriobacteriaceae, Eggerthellaceae, Enterobacteriaceae, Eubacteriaceae, and the like. ae), Lachnospiraceae, Lactobacillaceae, Leuconostocaceae, Odoribacteraceae, Oxalobacteraceae, Porphyromonadaceae, Prevotellaceae, Rikenellaceae, Ruminococcaceae, Streptococcaceae, and combinations thereof.

[0023] In certain embodiments, the cell compositions described herein, when stored at 4° C. for 6 or 12 months and then rehydrated, result in less than 3, 2, 1, or 0.5 log loss of colony forming units (CFU). In certain embodiments, the compositions remain substantially stable at 4° C. for about 2 years. In certain embodiments, the compositions, when rehydrated in an aqueous fluid, allow regrowth of one or more microorganisms within 5, 4, 3, or 2 hours of rehydration.

[0024] Additionally, the present disclosure provides pharmaceutical compositions comprising a cryopreserved composition of any of the cell-containing compositions described herein, optionally disposed in a capsule, tablet, or caplet. Additionally, the present disclosure provides food products or probiotics comprising a cryopreserved composition of any of the cell-containing compositions described herein, optionally disposed in a capsule, tablet, or caplet.

[0025] In another aspect, the present disclosure provides methods of modulating the gastrointestinal microbiome in a subject in need thereof, generally comprising administering to the subject an effective amount of any of the compositions, pharmaceutical compositions, or food products or probiotics described herein.

[0026] In another aspect, the disclosure provides a method of producing the cryopreservation composition described herein. The method generally includes: (a) combining an anaerobic microorganism with an excipient blend including an aqueous mixture including a viscosity modifier and an energy source for the microorganism to produce a cryopreservation mixture, the cryopreservation mixture having a viscosity in the range of about 5 cP to about 6,000 cP, an osmolality in the range of about 300 Osm / L to about 4,000 Osm / L, a pH in the range of about 4 to about 8, and optionally an oxidation-reduction potential in the range of -50 mV to -500 mV; and (b) lyophilizing the cryopreservation mixture to reduce the water content of the cryopreservation mixture to less than about 5% (w / w) to produce the cryopreservation composition. The microorganism can be in the form of a cell paste, for example, the cell paste can include the microorganism disposed in a cell culture medium. The method can further include packaging the cryopreservation composition into a capsule, tablet, or caplet. [Brief description of the drawings]

[0027] [Figure 1] 1 is a graph showing exemplary temperature and pressure profiles that can be used in the exemplary compositions disclosed herein for preserving anaerobic microorganisms. [Diagram 2] FIG. 1 is a bar graph showing the viability of microorganisms from the family Rikenellaceae, Bacteroidetaceae, Eubacteriumaceae, Lachnospiraceae, Lactobacillaceae, Leuconostocaceae, Oxalobacteraceae, and Streptococcus family after cryopreservation in (i) exemplary cryopreservation compositions of the present disclosure (ACP-001, ACP-002, and ACP-003), and (ii) commercially available cryopreservation compositions (CB-001 and CB-002, OPS DIagnostics catalog numbers LR2X-500-02 and MFDB-500-06), as described in Example 4. [Diagram 3]FIG. 13 is a graph showing optical density (OD600) as a function of ETF (time) for cryopreservation compositions of Bacteroidetes and Lachnospiraceae preserved using (i) an exemplary cryopreservation composition of the present disclosure and (ii) commercially available cryopreservation compositions (OPS DIagnostics catalog numbers LR2X-500-02 and MFDB-500-06), as described in Example 4. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0028] The present invention is based, in part, on the discovery of preservation compositions and techniques that promote long-term viability of anaerobic microorganisms and rapid recovery of the microorganisms when ingested by a subject so that the organisms can replicate as they traverse the subject's digestive tract. As generally described herein, the present disclosure provides compositions for preserving anaerobic microorganisms, including compositions for cryopreserving anaerobic microorganisms. The compositions generally include a viscosity modifier, an energy source for the anaerobic microorganisms to promote rapid recovery when administered to a subject, and an oxidation-reduction potential, e.g., about -50 mV to about -500 mV, to support the viability of the anaerobic organisms. Additionally, the compositions disclosed herein generally include one or more different types of anaerobic microorganisms, a viscosity modifier, an energy source for the microorganisms, and an oxidation-reduction potential, e.g., about -50 mV to about -500 mV, to support the viability of the anaerobic organisms.

[0029] definition To facilitate the understanding of this disclosure, several terms and phrases are defined below.

[0030] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. The abbreviations used herein have their conventional meaning in the chemical and biological arts.

[0031] Throughout the description, when compositions and kits are described as having, including, or comprising certain components, or when processes and methods are described as having, including, or comprising certain steps, it is further contemplated that there are compositions and kits of the present disclosure that consist essentially of or consist of the recited components, and that there are processes and methods according to the present disclosure that consist essentially of or consist of the recited processing steps.

[0032] Throughout this application, when an element or component is said to be included in and / or selected from a list of recited elements or components, it is understood that the element or component can be any of the recited elements or components, or the element or component can be selected from a group consisting of two or more of the recited elements or components.

[0033] Furthermore, it should be understood that the elements and / or features of the compositions or methods described herein can be combined in various ways without departing from the spirit and scope of the present disclosure, whether expressly or implicitly stated herein. For example, when a particular compound is mentioned, unless otherwise understood from the context, that compound can be used in various embodiments of the compositions of the present disclosure and / or in the methods of the present disclosure. In other words, within this application, the embodiments are described and depicted in a manner that allows a clear and concise application to be drawn, but it is intended and understood that the embodiments can be combined or separated in various ways without departing from the present teachings and disclosure. For example, it should be understood that all features described and depicted herein can be applied to all aspects of the present disclosure described and depicted herein.

[0034] The articles "a" and "an" are used in this disclosure, unless the content is inappropriate, to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element.

[0035] The term "and / or" is used in this disclosure to mean either "and" or "or," unless otherwise stated.

[0036] The phrase "at least one of" should be understood to include each of the listed items following the phrase and various combinations of two or more of the listed items individually, unless otherwise understood from the context and usage. The phrase "and / or" in the context of more than two listed items should be understood to have the same meaning, unless otherwise understood from the context.

[0037] Use of the terms "include," "includes," "including," "have," "has," "having," "contain," "contains," or "containing," including their grammatical equivalents, should generally be understood to be open-ended and open-ended, e.g., not excluding additional, unrecited elements or steps, unless otherwise specifically stated or understood from the context.

[0038] When the term "about" is used before a quantitative value, the present disclosure also includes the specific quantitative value itself, unless specifically stated otherwise. As used herein, the term "about" refers to a ±20% variation from the nominal value, unless otherwise specified or implied from internal usage.

[0039] At various places in this specification, variables or parameters are disclosed in groups or ranges. The description is specifically intended to include each and every individual subcombination of the members of such groups and ranges. For example, an integer in the range of 0 to 40 is specifically intended to disclose 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, and 40 individually, and an integer in the range of 1 to 20 is specifically intended to disclose 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20 individually.

[0040] The use of any and all examples, or exemplary language therein, such as "such as" or "including," is intended only to better illustrate the disclosure and does not impose limitations on the scope of the disclosure unless claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.

[0041] As a general matter, compositions specifying percentages are by weight unless otherwise specified. Further, if a variable is not accompanied by a definition, the previous definition of the variable will control.

[0042] As used herein, a "pharmaceutical composition" or "pharmaceutical formulation" refers to a combination of an active agent with inert or active excipients that make the composition or formulation particularly suitable for in vivo or ex vivo diagnostic or therapeutic uses.

[0043] "Pharmaceutically acceptable" means approved or approvable by a regulatory agency of the Federal or state government or a corresponding agency in a country outside the United States, or listed in the United States Pharmacopeia or other generally recognized pharmacopoeias, for use in animals, or more particularly, in humans.

[0044] As used herein, "pharmaceutical acceptable excipient" refers to a substance that supports the administration and / or absorption of active agents to a subject and can be included in the compositions of the present disclosure without causing significant adverse toxic effects to the patient.Non-limiting examples of pharmaceutical acceptable excipients include water, NaCl, normal saline such as phosphate buffered saline, emulsions (such as oil / water or water / oil emulsions), lactated Ringer's, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavorings, salt solutions (such as Ringer's solution), alcohol, oils, gelatin, carbohydrates such as lactose, amylose, or starch, fatty acid esters, hydroxymethylcellulose, polyvinylpyrrolidine, and colorants. Such preparations may be sterile and, if desired, mixed with auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring agents, and / or aromatic substances that do not deleteriously react with the compounds of the present disclosure. For examples of excipients, see Martin, REMINGTON'S PHARMACEUTICAL SCIENCES, 15th ed., Mack Publ. Co., Easton, PA (1975).

[0045] The terms "subject" and "patient" are used interchangeably herein. A "subject" or "patient" to which administration is contemplated includes, but is not limited to, humans (e.g., male or female of any age, e.g., pediatric subjects (e.g., infants, children, adolescents) or adult subjects (e.g., young adults, middle-aged adults, or geriatrics)) and / or non-human animals, e.g., mammals such as primates (e.g., cynomolgus monkeys, rhesus monkeys), cattle, pigs, horses, sheep, goats, rodents, cats, and / or dogs. In certain embodiments, the subject is a human. In certain embodiments, the subject is a non-human animal.

[0046] As used herein, "administering" refers to oral administration, administration as a suppository, topical contact, intravenous administration, parenteral administration, intraperitoneal administration, intramuscular administration, intralesional administration, intrathecal administration, intracranial administration, intranasal administration, or subcutaneous administration, or implantation of a sustained release device, such as a mini-osmotic pump, into a subject. Administration can be by any route, including parenteral and transmucosal (e.g., buccal, sublingual, palatal, gingival, nasal, vaginal, rectal, or transdermal). Parenteral administration includes, for example, intravenous, intramuscular, intraarterial, intradermal, subcutaneous, intraperitoneal, intraventricular, and intracranial. Other modes of delivery include, but are not limited to, the use of liposomal formulations, intravenous infusion, transdermal patches, and the like. In certain embodiments, a preserved composition (e.g., a cryopreserved composition) as described herein is orally administered to a subject. "Co-administering" refers to administering a composition as described herein simultaneously with, immediately before, or immediately after administration of one or more additional therapies. The preserved compositions (e.g., cryopreserved compositions) described herein can be administered alone or simultaneously to a subject. Simultaneous administration is meant to include simultaneous or sequential administration of the compositions, individually or in combination (multiple compositions or agents). Thus, the preparations can also be combined with other active substances / compositions as desired.

[0047] The terms "disease," "disorder," and "condition" are used interchangeably herein.

[0048] Unless otherwise specified, the terms "treat," "treating," and "treatment" as used herein contemplate an action occurring during the course of suffering from a specified disease, disorder, or condition that reduces the severity of the disease, disorder, or condition, or that slows or retards the progression of the disease, disorder, or condition (e.g., "therapeutic treatment").

[0049] In general, an "effective amount" of a composition (e.g., a preserved or cryopreserved composition described herein) refers to an amount sufficient to induce a desired biological response, e.g., to modulate the gastrointestinal microbiome of a subject. As will be appreciated by those skilled in the art, an effective amount of a composition of the present disclosure (e.g., a preserved or cryopreserved composition described herein) can vary depending on factors such as, for example, the desired biological endpoint, the residence time and viability of the composition, the disease being treated, and the age, weight, health, and condition of the subject.

[0050] The term "anaerobic microorganisms" as used herein is understood to mean microorganisms that can remain viable and / or grow in an oxygen-depleted environment. The term anaerobic microorganisms includes obligate anaerobes, facultative anaerobes, and aerotolerant microorganisms. In certain embodiments, anaerobic microorganisms can remain viable, grow, and / or replicate in an environment with an oxidation-reduction potential of about +300 mV to -100 mV for facultative anaerobes and +100 mV to about -500 mV for obligate anaerobes.

[0051] I. Compositions for Preserving Anaerobic Microorganisms In one aspect, provided herein is a composition for preserving anaerobic microorganisms, generally comprising a viscosity modifier and an energy source for the microorganisms. In certain embodiments, the composition for preserving anaerobic microorganisms described herein is a cryopreservation composition for cryopreserving anaerobic microorganisms.

[0052] In certain embodiments, the composition, when solvated in an aqueous solvent, has a viscosity in the range of about 5 cP to about 6,000 cP. Alternatively or additionally, the composition, when solvated in an aqueous solvent, has an osmolality in the range of about 300 Osm / L to about 4,000 Osm / L, a redox potential in the range of about -50 mV to about -500 mV, and / or a pH in the range of about 4 to about 8.

[0053] In certain embodiments, a storage (e.g., cryopreservation) composition for preserving anaerobic microorganisms comprises a viscosity modifier and an energy source for the microorganism, and optionally (i) has a redox potential in the range of about -50 mV to about -500 mV, and (ii) when solvated in an aqueous solvent, has a viscosity in the range of about 5 cP to about 6,000 cP, an osmolality in the range of about 300 Osm / L to about 4,000 Osm / L, a redox potential in the range of about -50 mV to about -500 mV, and a pH in the range of about 4 to about 8.

[0054] In certain embodiments, a cryopreservation composition for cryopreserving anaerobic microorganisms comprises a viscosity modifier, an energy source for the microorganism, and a redox potential in the range of about -50 mV to about -500 mV, and when solvated in an aqueous solvent has a viscosity in the range of about 5 cP to about 6,000 cP, an osmolality in the range of about 300 Osm / L to about 4,000 Osm / L, a redox potential in the range of about -50 mV to about -500 mV, and a pH in the range of about 4 to about 8.

[0055] Viscosity and Viscosity Modifiers The viscosity is selected to maintain the viability of the preserved microorganism or microorganisms.

[0056] In certain embodiments, the composition, when solvated in an aqueous solvent, has a molecular weight of about 5 cP to about 6,000 cP, about 50 cP to about 6,000 cP, about 100 cP to about 6,000 cP, about 200 cP to about 6,000 cP, about 300 cP to about 6,000 cP, about 400 cP to about 6,000 cP, about 500 cP to about 6,000 cP, about 1,000 cP to about 6,000 cP, about 1,500 cP to about 6,000 cP, about 2,000 cP to about 6,000 cP, about 2,500 cP to about 6,000 cP, about 3,000 cP to about 6,000 cP, about 3,500 cP to about 6,000 cP, 0cP, approximately 4,000cP to approximately 6,000cP, approximately 4,500cP to approximately 6,000cP, approximately 5,000cP to approximately 6,000cP, approximately 5, 500cP~Approx. 6,000cP, Approx. 5cP~Approx. 5,500cP, Approx. 5cP~Approx. 5,000cP, Approx. 5cP~Approx. 5,000cP, Approx. 5cP~ Approx. 4,500cP, Approx. 5cP ~ Approx. 4,000cP, Approx. 5cP ~ Approx. 3,500cP, Approx. 5cP ~ Approx. 3,000cP, Approx. 5cP ~ Approx. 2,500 cP, about 5 cP to about 2,000 cP, about 5 cP to about 1,500 cP, about 5 cP to about 1,000 cP, about 5 cP to about 500 cP, about 5 cP to Approximately 400cP, approximately 5cP to approximately 300cP, approximately 5cP to approximately 200cP, approximately 5cP to approximately 100cP, approximately 5cP to approximately 50cP, approximately 50cP to approximately 5,500cP, about 50cP to about 5,000cP, about 50cP to about 4,500cP, about 50cP to about 4,000cP, about 50cP to about 3, 500cP, about 50cP to about 3,000cP, about 50cP to about 2,500cP, about 50cP to about 2,000cP, about 50cP to about 1,50 0cP, about 50cP to about 1,000cP, about 50cP to about 500cP, about 50cP to about 400cP, about 50cP to about 300cP, about 50c P ~ about 200cP, about 50cP - about 100cP, about 100cP - about 5,500cP, about 100cP - about 5,000cP, about 100cP - about 4,500cP, about 100cP to about 4,000cP, about 100cP to about 3,500cP, about 100cP to about 3,000cP, about 100cP ~2,500cP, 100cP~2,000cP, 100cP~1,500cP, 100cP~1,000cP, 100 cP ~ approx. 500cP, approx. 100cP ~ approx. 400cP, approx. 100cP ~ approx. 300cP, approx. 100cP ~ approx. 200cP, approx. 200cP ~ approx. 5,500 cP, from about 200 cP to about 5,000 cP, from about 200 cP to about 4,500 cP, from about 200 cP to about 4,000 cP, from about 200 cP to about 3,500 cP, from about 200 cP to about 3,000 cP, from about 200 cP to about 2,500 cP, from about 200 cP to about 2,000 cP, from about 200 cP to about 1,500 cP, from about 200 cP to about 1,000 cP, from about 200 cP to about 500 cP, from about 200 cP to about 400 cP, from about 200 cP to about 300 cP, from about 300 cP to about 5,500 cP, from about 300 cP to about 5,000 cP, from about 300 cP to about 4,500 cP, from about 300 cP to about 4,000 cP, from about 300 cP to about 3,500 cP, from about 300 cP to about 3,000 cP, from about 300 cP to about 2,500 cP, from about 300 cP to about 2,000 cP, from about 300 cP to about 1,500 cP, from about 300 cP to about 1,000 cP, from about 300 cP to about 500 cP, from about 300 cP to about 400 cP, from about 400 cP to about 5,500 cP, from about 400 cP to about 5,000 cP, from about 400 cP to about 4,500 cP, from about 400 cP to about 4,000 cP, from about 400 cP to about 3,500 cP, from about 400 cP to about 3,000 cP, from about 400 cP to about 2,500 cP, from about 400 cP to about 2,000 cP, from about 400 cP to about 1,500 cP, from about 400 cP to about 1,000 cP, from about 400 cP to about 500 cP, from about 500 cP to about 5,500 cP, from about 500 cP to about 5,000 cP, from about 500 cP to about 4,500 cP, from about 500 cP to about 4,000 cP, from about 500 cP to about 3,500 cP, from about 500 cP to about 3,000 cP, from about 500 cP to about 2,500 cP, from about 500 cP to about 2,000 cP, from about 500 cP to about 1,500 cP, from about 500 cP to about 1,000 cP, from about 1,000 cP to about 5,500 cP, from about 1,000 cP to about 5,000 cP, from about 1,000 cP to about 4,500 cP, from about 1,000 cP to about 4,000 cP, from about 1,000 cP to about 3,500 cP, from about 1,000 cP to about 3,000 cP, from about 1,000 cP to about 2,500 cP, from about 1,000 cP to about 2,000 cP, from about 1,000 cP to about 1,500 cP, from about 1,500 cP to about 5,500 cP, from about 1,500 cP to about 5,000 cP, from about 1,500 cP to about 4,500 cP, from about 1,500 cP to about 4,000 cP, from about 1,500 cP to about 3,500 cP, from about 1,500 cP to about 3,000 cP, from about 1,500 cP to about 2,500cP, approximately 1,500cP to approximately 2,000cP, approximately 2,000cP to approximately 5,500cP, approximately 2,000cP to approximately 5,000cP, approximately 2,000c P ~ about 4,500cP, about 2,000cP - about 4,000cP, about 2,000cP - about 3,500cP, about 2,000cP - about 3,000cP, about 2 ,000cP ~ approx. 2,500cP, approx. 2,500cP ~ approx. 5,500cP, approx. 2,500cP ~ approx. 5,000cP, approx. 2,500cP ~ approx. 4,500 cP, approximately 2,500cP to approximately 4,000cP, approximately 2,500cP to approximately 3,500cP, approximately 2,500cP to approximately 3,000cP, approximately 3,000cP to approximately 5 ,500cP, approx. 3,000cP ~ approx. 5,000cP, approx. 3,000cP ~ approx. 4,500cP, approx. 3,000cP ~ approx. 4,000cP, approx. 3,000 cP ~ approx. 3,500cP, approx. 3,500cP ~ approx. 5,500cP, approx. 3,500cP ~ approx. 5,000cP, approx. 3,500cP ~ approx. 4,500cP, approx. It has a viscosity in the range of 3,500 cP to about 4,000 cP, about 4,000 cP to about 5,500 cP, about 4,000 cP to about 5,000 cP, about 4,000 cP to about 4,500 cP, about 4,500 cP to about 5,500 cP, about 4,500 cP to about 5,000 cP, about 5,000 cP to about 5,500 cP.

[0057] In certain embodiments, the viscosity is in the range of about 50 cP to about 4,000 cP, about 100 cP to about 2,000 cP, or about 100 cP to 1,000 cP when solvated in an aqueous solvent.

[0058] In certain embodiments, the composition, when solvated in an aqueous solvent, has a viscosity of about 5 cP, about 10 cP, about 15 cP, about 20 cP, about 25 cP, about 30 cP, about 35 cP, about 40 cP, about 45 cP, about 50 cP, about 55 cP, about 60 cP, about 65 cP, about 70 cP, about 75 cP, about 80 cP, about 85 cP, about 90 cP, about 95 cP, about 100 cP, about 110 cP, about 120 cP, about 130 cP, about 140 cP, about 150 cP, about 160 cP, about 170 cP, about 180 cP, about The viscosity is about 190 cP, about 200 cP, about 225 cP, about 250 cP, about 275 cP, about 300 cP, about 350 cP, about 400 cP, about 450 cP, about 500 cP, about 600 cP, about 700 cP, about 800 cP, about 900 cP, about 1,000 cP, about 1,250 cP, about 1,500 cP, about 2,000 cP, about 2,500 cP, about 3,000 cP, about 3,500 cP, about 4,000 cP, about 4,500 cP, about 5,000 cP, about 5,500 cP, or about 6,000 cP.

[0059] In certain embodiments, the composition, when solvated in an aqueous solvent, is from about 0.01% (w / w) to about 0.4% (w / w), from about 0.05% (w / w) to about 0.4% (w / w), from about 0.1% (w / w) to about 0.4% (w / w), from about 0.15% (w / w) to about 0.4% (w / w), from about 0.2% (w / w) to about 0.4% (w / w), from about 0.25% (w / w) to about 0.4% (w / w), from about 0.3% (w / w) to about 0.4% (w / w), from about 0.35% (w / w) to about 0.4% (w / w), from about 0.0 1% (w / w) to about 0.35% (w / w), about 0.01% (w / w) to about 0.3% (w / w), about 0.01% (w / w) to about 0.25% (w / w), about 0.01% (w / w) to about 0.2% (w / w), about 0.01% (w / w) to about 0.15% (w / w), about 0.01% (w / w) to about 0.1% (w / w), about 0.01% (w / w) to about 0.05% (w / w), about 0.05% (w / w) to about 0.35% (w / w), about 0.05% (w / w) to about 0.3% (w / w), about 0.05% (w / w) to About 0.25% (w / w), about 0.05% (w / w) to about 0.2% (w / w), about 0.05% (w / w) to about 0.15% (w / w), about 0.05% (w / w) to about 0.1% (w / w), about 0.1% (w / w) to about 0.35% (w / w), about 0.1% (w / w) to about 0.3% (w / w), about 0.1% (w / w) to about 0.25% (w / w), about 0.1% (w / w) to about 0.2% (w / w), about 0.1% (w / w) to about 0.15% (w / w), about 0.15% (w / w) to about 0.35% (w / w), The viscosity modifier may be about 0.15% (w / w) to about 0.3% (w / w), about 0.15% (w / w) to about 0.25% (w / w), about 0.15% (w / w) to about 0.2% (w / w), about 0.2% (w / w) to about 0.35% (w / w), about 0.2% (w / w) to about 0.3% (w / w), about 0.2% (w / w) to about 0.25% (w / w), about 0.25% (w / w) to about 0.35% (w / w), about 0.25% (w / w) to about 0.35% (w / w), or about 0.3% (w / w) to about 0.35% (w / w). In certain embodiments, the composition comprises from about 0.01% (w / w) to about 0.4% (w / w) of a viscosity modifier when solvated in an aqueous solvent.

[0060] In certain embodiments, the composition, when solvated in an aqueous solvent, is about 0.01% (w / w), about 0.02% (w / w), about 0.03% (w / w), about 0.04% (w / w), about 0.05% (w / w), about 0.06% (w / w), about 0.07% (w / w), about 0.08% (w / w), about 0.09% (w / w), about 0.1% (w / w), about 0.11% (w / w), about 0.12% (w / w), about 0.13% (w / w), about 0.14% (w / w), about 0.15% (w / w), about 0.16% (w / w), about 0.17% (w / w), about 0.18% (w / w), about 0.19% (w / w), Approximately 0.2% (w / w), approximately 0.21% (w / w), approximately 0.22% (w / w), approximately 0.23% (w / w), approximately 0.24% (w / w), approximately 0.25% (w / w), approximately 0.26% (w / w), approximately 0.27% (w / w), approximately 0.28% (w / w), approximately 0.29% (w / w), approximately 0.3% (w / w), About 0.31% (w / w), about 0.32% (w / w), about 0.33% (w / w), about 0.34% (w / w), about 0.35% (w / w), about 0.36% (w / w), about 0.37% (w / w), about 0.38% (w / w), about 0.39% (w / w), or about 0.4% (w / w) of a viscosity modifier.

[0061] In certain embodiments, the viscosity modifier is a polymer composition. In certain embodiments, the polymer composition comprises alginate, acacia, alginic acid, agar, aramic acid, bentonite, carbomer, carrageenan, cellulose, dextrin, gelatin, gellan gum, guar gum, carboxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, methylcellulose, mucin, pectin, starch, xanthan gum, or a combination thereof. In certain embodiments, the polymer composition comprises an alginate, such as sodium alginate.

[0062] In certain embodiments, sodium alginate has a viscosity of about 250 cP to about 14,000 cP, about 500 cP to about 14,000 cP, about 750 cP to about 14,000 cP, about 1,000 cP to about 14,000 cP, about 2,000 cP to about 14,000 cP, about 3,000 cP to about 14,000 cP, about 4,000 cP to about 14,000 cP, about 5,000 cP to about 14,000 cP, about 6,000 cP to about 14,000 cP, about 7,000 cP to about 14,000 cP, about 8,000 cP to about 14,000 cP, about 9,000 cP to about 14,000 cP, or about 10 ,000cP ~ approx. 14,000cP, approx. 11,000cP ~ approx. 14,000cP, approx. 12,000cP ~ approx. 14,000cP, approx. 13,0 00cP ~ approx. 14,000cP, approx. 250cP ~ approx. 13,000cP, approx. 250cP ~ approx. 12,000cP, approx. 250cP ~ approx. 11,000 cP, about 250cP to about 10,000cP, about 250cP to about 9,000cP, about 250cP to about 8,000cP, about 250cP to about 7, 000cP, about 250cP to about 6,000cP, about 250cP to about 5,000cP, about 250cP to about 4,000cP, about 250cP to about 3 ,000cP, about 250cP to about 2,000cP, about 250cP to about 1,000cP, about 250cP to about 750cP, about 250cP to about 5 00cP, approx. 500cP ~ approx. 13,000cP, approx. 500cP ~ approx. 12,000cP, approx. 500cP ~ approx. 11,000cP, approx. 500cP ~10,000cP, 500cP~9,000cP, 500cP~8,000cP, 500cP~7,000cP, 50 0cP ~ approx. 6,000cP, approx. 500cP ~ approx. 5,000cP, approx. 500cP ~ approx. 4,000cP, approx. 500cP ~ approx. 3,000cP, approx. 5 00cP~Approx. 2,000cP, Approx. 500cP~Approx. 1,000cP, Approx. 500cP~Approx. 750cP, Approx. 750cP~Approx. 13,000cP, Approx. 750cP~Approx. 12,000cP, Approx. 750cP~Approx. 11,000cP, Approx. 750cP~Approx. 10,000cP, Approx. 750cP~Approx. 9,000 cP, about 750cP to about 8,000cP, about 750cP to about 7,000cP, about 750cP to about 6,000cP, about 750cP to about 5,0 00cP, about 750cP to about 4,000cP, about 750cP to about 3,000cP, about 750cP to about 2,000cP, about 750cP to about 1,000 cP, from about 1,000 cP to about 13,000 cP, from about 1,000 cP to about 12,000 cP, from about 1,000 cP to about 11,000 cP, from about 1,000 cP to about 10,000 cP, from about 1,000 cP to about 9,000 cP, from about 1,000 cP to about 8,000 cP, from about 1,000 cP to about 7,000 cP, from about 1,000 cP to about 6,000 cP, from about 1,000 cP to about 5,000 cP, from about 1,000 cP to about 4,000 cP, from about 1,000 cP to about 3,000 cP, from about 1,000 cP to about 2,000 cP, from about 2,000 cP to about 13,000 cP, from about 2,000 cP to about 12,000 cP, from about 2,000 cP to about 11,000 cP, from about 2,000 cP to about 10,000 cP, from about 2,000 cP to about 9,000 cP, from about 2,000 cP to about 8,000 cP, from about 2,000 cP to about 7,000 cP, from about 2,000 cP to about 6,000 cP, from about 2,000 cP to about 5,000 cP, from about 2,000 cP to about 4,000 cP, from about 2,000 cP to about 3,000 cP, from about 3,000 cP to about 13,000 cP, from about 3,000 cP to about 12,000 cP, from about 3,000 cP to about 11,000 cP, from about 3,000 cP to about 10,000 cP, from about 3,000 cP to about 9,000 cP, from about 3,000 cP to about 8,000 cP, from about 3,000 cP to about 7,000 cP, from about 3,000 cP to about 6,000 cP, from about 3,000 cP to about 5,000 cP, from about 3,000 cP to about 4,000 cP, from about 4,000 cP to about 13,000 cP, from about 4,000 cP to about 12,000 cP, from about 4,000 cP to about 11,000 cP, from about 4,000 cP to about 10,000 cP, from about 4,000 cP to about 9,000 cP, from about 4,000 cP to about 8,000 cP, from about 4,000 cP to about 7,000 cP, from about 4,000 cP to about 6,000 cP, from about 4,000 cP to about 5,000 cP, from about 5,000 cP to about 13,000 cP, from about 5,000 cP to about 12,000 cP, from about 5,000 cP to about 11,000 cP, from about 5,000 cP to about 10,000 cP, from about 5,000 cP to about 9,000 cP, from about 5,000 cP to about 8,000 cP, from about 5,000 cP to about 7,000 cP, from about 5,000 cP to about 6,000 cP, from about 6,000 cP to about 13,000 cP, from about 6,000 cP to about 12,000 cP, from about 6,000 cP to about 11,000 cP, from about 6,000 cP to about 10,000cP, approx. 6,000cP ~ approx. 9,000cP, approx. 6,000cP ~ approx. 8,000cP, approx. 6,000cP ~ approx. 7,000cP , about 7,000cP to about 13,000cP, about 7,000cP to about 12,000cP, about 7,000cP to about 11,000cP, about 7,000cP~approx. 10,000cP, approx. 7,000cP~approx. 9,000cP, approx. 7,000cP~approx. 8,000cP, approx. 8,00 0cP ~ approx. 13,000cP, approx. 8,000cP ~ approx. 12,000cP, approx. 8,000cP ~ approx. 11,000cP, approx. 8,000cP to about 10,000 cP, about 8,000 cP to about 9,000 cP, about 9,000 cP to about 13,000 cP, about 9,000 cP to about 12,000 cP, about 9,000 cP to about 11,000 cP, about 9,000 cP to about 10,000 cP, about 10,000 cP to about 13,000 cP, about 10,000 cP to about 12,000 cP, about 10,000 cP to about 11,000 cP, about 11,000 cP to about 13,000 cP, about 11,000 cP to about 12,000 cP, or about 12,000 cP to about 13,000 cP. In certain embodiments, the sodium alginate has a viscosity range of about 250 cP to about 14,000 cP.

[0063] Osmolality and Osmolality Adjusters The osmolality is selected to maintain the viability of the microorganism or microorganisms being stored.

[0064] In certain embodiments, the composition, when solvated in an aqueous solvent, has a molecular weight of about 300 Osm / L to about 4,000 Osm / L, about 400 Osm / L to about 4,000 Osm / L, about 500 Osm / L to about 4,000 Osm / L, about 600 Osm / L to about 4,000 Osm / L, about 700 Osm / L to about 4,000 Osm / L, about 800 Osm / L to about 4,000 Osm / L, about 900 Osm / L to about 4,000 Osm / L, about 1,000 Osm / L to about 4,000 Osm / L, about 1,500 Osm / L to about 4,000 Osm / L, about 2,000 Osm / L, or about 3,000 Osm / L. / L ~ approx. 4,000Osm / L, approx. 2,500Osm / L ~ approx. 4,000Osm / L, approx. 3,000Osm / L ~ approx. 4,000Osm / L, about 3,500Osm / L~about 4,000Osm / L, about 300Osm / L~about 3,500Osm / L, about 300Osm / L~about 3,0 00Osm / L, about 300Osm / L to about 2,500Osm / L, about 300Osm / L to about 2,000Osm / L, about 300Osm / L ~Approx. 1,500Osm / L, Approx. 300Osm / L ~ Approx. 1,000Osm / L, Approx. 300Osm / L ~ Approx. 900Osm / L, Approx. 300Osm / L ~ Approx. 800Osm / L, Approx. 300Osm / L ~ Approx. 700Osm / L, Approx. 300Osm / L ~ Approx. 600Osm / L, Approx. 300Osm / L ~ approx. 500Osm / L, approx. 300Osm / L ~ approx. 400Osm / L, approx. 400Osm / L ~ approx. 3,500Osm / L, approx. 400Osm / L ~ approx. 3,000Osm / L, approx. 400Osm / L ~ approx. 2,500Osm / L, approx. 400Osm / L ~ approx. 2,500Osm / L, approx. 40 0Osm / L~Approx. 2,000Osm / L, Approx. 400Osm / L~Approx. 2,000Osm / L, Approx. 400Osm / L~Approx. 1,500Osm / L , about 400Osm / L~about 1,000Osm / L, about 400Osm / L~about 900Osm / L, about 400Osm / L~about 800Osm / L, about 400Osm / L to about 700Osm / L, about 400Osm / L to about 600Osm / L, about 400Osm / L to about 500Osm / L, Approx. 500Osm / L ~ Approx. 3,500Osm / L, Approx. 500Osm / L ~ Approx. 3,000Osm / L, Approx. 500Osm / L ~ Approx. 2,500O sm / L, about 500Osm / L to about 2,000Osm / L, about 500Osm / L to about 1,500Osm / L, about 500Osm / L to about 1,000 Osm / L, from about 500 Osm / L to about 900 Osm / L, from about 500 Osm / L to about 800 Osm / L, from about 500 Osm / L to about 700 Osm / L, from about 500 Osm / L to about 600 Osm / L, from about 600 Osm / L to about 3,500 Osm / L, from about 600 Osm / L to about 3,000 Osm / L, from about 600 Osm / L to about 2,500 Osm / L, from about 600 Osm / L to about 2,000 Osm / L, from about 600 Osm / L to about 1,500 Osm / L, from about 600 Osm / L to about 1,000 Osm / L, from about 600 Osm / L to about 900 Osm / L, from about 600 Osm / L to about 800 Osm / L, from about 600 Osm / L to about 700 Osm / L, from about 700 Osm / L to about 3,500 Osm / L, from about 700 Osm / L to about 3,000 Osm / L, from about 700 Osm / L to about 2,500 Osm / L, from about 700 Osm / L to about 2,000 Osm / L, from about 700 Osm / L to about 1,500 Osm / L, from about 700 Osm / L to about 1,000 Osm / L, from about 700 Osm / L to about 900 Osm / L, from about 700 Osm / L to about 800 Osm / L, from about 800 Osm / L to about 3,500 Osm / L, from about 800 Osm / L to about 3,000 Osm / L, from about 800 Osm / L to about 2,500 Osm / L, from about 800 Osm / L to about 2,000 Osm / L, from about 800 Osm / L to about 1,500 Osm / L, from about 800 Osm / L to about 1,000 Osm / L, from about 800 Osm / L to about 900 Osm / L, from about 900 Osm / L to about 3,500 Osm / L, from about 900 Osm / L to about 3,000 Osm / L, from about 900 Osm / L to about 2,500 Osm / L, from about 900 Osm / L to about 2,000 Osm / L, from about 900 Osm / L to about 1,500 Osm / L, from about 900 Osm / L to about 1,000 Osm / L, from about 1,000 Osm / L to about 3,500 Osm / L, from about 1,000 Osm / L to about 3,000 Osm / L, from about 1,000 Osm / L to about 2,500 Osm / L, from about 1,000 Osm / L to about 2,000 Osm / L, from about 1,000 Osm / L to about 1,500 Osm / L, from about 1,500 Osm / L to about 3,500 Osm / L, from about 1,500 Osm / L to about 3,000 Osm / L, from about 1,500 Osm / L to about 2,500 Osm / L, from about 1,500 Osm / L to about 2,000 Osm / L, from about 2,000 Osm / L to about 3,500 Osm / L, from about 2,000 Osm / L to about 3,The osmolality ranges from about 2,000 Osm / L to about 2,500 Osm / L, about 2,500 Osm / L to about 3,500 Osm / L, about 2,500 Osm / L to about 3,000 Osm / L, or about 3,000 Osm / L to about 3,500 Osm / L.

[0065] In certain embodiments, the composition, when solvated in an aqueous solvent, has an osmolality in the range of about 300 Osm / L to about 3,000 Osm / L, about 400 Osm / L to about 3,000 Osm / L, or about 500 Osm / L to about 3,000 Osm / L.

[0066] In certain embodiments, the composition, when solvated in an aqueous solvent, has an osmolality of about 300 Osm / L, about 350 Osm / L, about 400 Osm / L, about 450 Osm / L, about 500 Osm / L, about 600 Osm / L, about 700 Osm / L, about 800 Osm / L, about 900 Osm / L, about 1,000 Osm / L, about 1,250 Osm / L, about 1,500 Osm / L, about 1,750 Osm / L, about 2,000 Osm / L, about 2,250 Osm / L, about 2,500 Osm / L, about 2,750 Osm / L, about 3,000 Osm / L, about 3,500 Osm / L, or about 4,000 Osm / L.

[0067] Osmolality can be adjusted using various agents, such as salts, sugars, and polymers. Exemplary salts include, but are not limited to, NaCl, KCl, MgCl 2 , CaCl 2 , Na 2 HPO 4 , NaH 2 PO 4 , K 2 HPO 4 , K.H. 2 PO 4Exemplary sugars include, but are not limited to, monosaccharides and disaccharides, such as glucose, sucrose, trehalose. Exemplary polymers include, but are not limited to, alginate, acacia, alginic acid, agar, aramic acid, bentonite, carbomer, carrageenan, cellulose, dextrin, gelatin, gellan gum, guar gum, carboxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, methylcellulose, mucin, pectin, starch, xanthan gum, or combinations thereof.

[0068] pH The pH is selected to maintain the viability of the one or more microorganisms being preserved. In certain embodiments, the composition, when solvated in an aqueous solvent, is from about 4 to about 8, from about 4.5 to about 8, from about 5 to about 8, from about 5.5 to about 8, from about 6 to about 8, from about 6.5 to about 8, from about 7 to about 8, from about 7.5 to about 8, from about 4 to about 7.5, from about 4 to about 7, from about 4 to about 6.5, from about 4 to about 6, from about 4 to about 5.5, from about 4 to about 5, from about 4 to about 4.5, from about 4.5 to about 7.5, from about 4.5 to about 7, from about 4.5 to about 5. The pH range is about 6.5, about 4.5 to about 6, about 4.5 to about 5.5, about 4.5 to about 5, about 5 to about 7.5, about 5 to about 7, about 5 to about 6.5, about 5 to about 6, about 5 to about 5.5, about 5.5 to about 7.5, about 5.5 to about 7, about 5.5 to about 6.5, about 5.5 to about 6, about 6 to about 7.5, about 6 to about 7, about 6 to about 6.5, about 6.5 to about 7.5, about 6.5 to about 7, or about 7 to about 7.5.

[0069] In certain embodiments, the composition, when solvated in an aqueous solvent, has a pH in the range of about 4 to about 8, about 5 to about 8, about 4 to about 7.5, about 5 to about 7.5, or about 5.5 to about 7.5.

[0070] In certain embodiments, the composition has a pH of about 4, about 4.1, about 4.2, about 4.3, about 4.4, about 4.5, about 4.6, about 4.7, about 4.8, about 4.9, about 5, about 5.1, about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, about 6, about 6.1, about 6.2, about 6.3, about 6.4, about 6.5, about 6.6, about 6.7, about 6.8, about 6.9, about 7, about 7.1, about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7, about 7.8, about 7.9, or about 8 when solvated in an aqueous solvent.

[0071] Energy source for microorganisms In certain embodiments, the energy source is a cell culture medium that allows the growth of the microorganism. It will be understood that the culture medium may vary depending on the anaerobic microorganism of interest and may therefore be tailored to a given microorganism or group of microorganisms. Exemplary culture media may contain one or more of the following components, including, but not limited to, carbon sources (e.g., sugars), nitrogen sources (e.g., ammonia-rich components (e.g., yeast extract, peptone) and amino acids), vitamins, minerals (e.g., phosphate, acetate), buffer components (e.g., sodium phosphate or potassium phosphate buffer), trace metals, fatty acids, and oxygen scavengers (e.g., cysteine, sodium dithionite, sodium sulfite, thioglycolate, ferric sulfide, dithiothreitol).

[0072] In certain embodiments, the composition, when solvated in an aqueous solvent, is from about 3% (w / w) to about 15% (w / w), from about 5% (w / w) to about 15% (w / w), from about 7% (w / w) to about 15% (w / w), from about 9% (w / w) to about 15% (w / w), from about 11% (w / w) to about 15% (w / w), from about 13% (w / w) to about 15% (w / w), from about 3% (w / w) to about 13% (w / w), from about 3% (w / w) to about 11% (w / w), from about 3% (w / w) to about 9% (w / w), from about 3% (w / w) to about 7% (w / w),

[0043] The energy source may comprise about 3% (w / w) to about 5% (w / w), about 5% (w / w) to about 13% (w / w), about 5% (w / w) to about 11% (w / w), about 5% (w / w) to about 9% (w / w), about 5% (w / w) to about 7% (w / w), about 7% (w / w) to about 13% (w / w), about 7% (w / w) to about 11% (w / w), about 7% (w / w) to about 9% (w / w), about 9% (w / w) to about 13% (w / w), about 9% (w / w) to about 11% (w / w), or about 11% (w / w) to about 13% (w / w). In certain embodiments, the composition comprises from about 3% (w / w) to about 15% (w / w) of the energy source when solvated in an aqueous solvent.

[0073] In certain embodiments, the composition comprises about 3% (w / w), about 4% (w / w), about 5% (w / w), about 6% (w / w), about 7% (w / w), about 8% (w / w), about 9% (w / w), about 10% (w / w), about 11% (w / w), about 12% (w / w), about 13% (w / w), about 14% (w / w), or about 15% (w / w) of the energy source.

[0074] In certain embodiments, the energy source is a monosaccharide, a disaccharide, an oligosaccharide, or a combination thereof. In certain embodiments, the monosaccharide is selected from the group consisting of glucose, fructose, xylose, and combinations thereof. In certain embodiments, the disaccharide is trehalose, sucrose, or a combination thereof. In certain embodiments, the oligosaccharide is an inulin derivative, such as a fructooligosaccharide, such as raftilose. In certain embodiments, the oligosaccharide is maltodextrin.

[0075] In certain embodiments, the energy source comprises raftilose and maltodextrin.

[0076] In certain embodiments, the composition, when solvated in an aqueous solvent, is from about 0.5% (w / w) to about 5% (w / w), from about 1% (w / w) to about 5% (w / w), from about 1.5% (w / w) to about 5% (w / w), from about 2% (w / w) to about 5% (w / w), from about 2.5% (w / w) to about 5% (w / w), from about 3% (w / w) to about 5% (w / w), from about 3.5% (w / w) to about 5% (w / w), from about 4% (w / w) to about 5% (w / w), from about 4.5% (w / w) to about 5% (w / w), from about 0.5% (w / w) to about 4.5% (w / w), or from about 0.5% (w / w) to about 4.5% (w / w). (w / w) to about 4% (w / w), about 0.5% (w / w) to about 3.5% (w / w), about 0.5% (w / w) to about 3% (w / w), about 0.5% (w / w) to about 2.5% (w / w), about 0.5% (w / w) to about 2% (w / w), about 0.5% (w / w) to about 1.5% (w / w), about 0.5% (w / w) to about 1% (w / w), about 1% (w / w) to about 4.5% (w / w), about 1% (w / w) to about 4% (w / w), about 1% (w / w) to about 3.5% (w / w), about 1% (w / w) to about 3% (w / w), about 1% (w / w) to about 2.5% (w / w), about 1% (w / w) to about 2% (w / w), about 1% (w / w) to about 1.5% (w / w), about 1.5% (w / w) to about 4.5% (w / w), about 1.5% (w / w) to about 4% (w / w), about 1.5% (w / w) to about 3.5% (w / w), about 1.5% (w / w) to about 3% (w / w), about 1.5% (w / w) to about 2.5% (w / w), about 1.5% (w / w) to about 2% (w / w), about 2% (w / w) to about 4.5% (w / w), about 2% (w / w) to about 4% (w / w), about 2% (w / w) to about 3.5% (w / w), about 2% (w / w) to about 3% (w / w), The composition contains about 2% (w / w) to about 2.5% (w / w), about 2.5% (w / w) to about 4.5% (w / w), about 2.5% (w / w) to about 4% (w / w), about 2.5% (w / w) to about 3.5% (w / w), about 2.5% (w / w) to about 3% (w / w), about 3% (w / w) to about 4.5% (w / w), about 3% (w / w) to about 4% (w / w), about 3% (w / w) to about 3.5% (w / w), about 3.5% (w / w) to about 4.5% (w / w), about 3.5% (w / w) to about 4% (w / w), or about 4% (w / w) to about 4.5% (w / w).In certain embodiments, the composition comprises from about 0.5% (w / w) to about 5% (w / w) of raftilose when solvated in an aqueous solvent.

[0077] In certain embodiments, the composition contains about 0.5% (w / w), about 1% (w / w), about 1.5% (w / w), about 2% (w / w), about 3% (w / w), about 3.5% (w / w), about 4% (w / w), about 4.5% (w / w), or about 5% (w / w) of raftilose when solvated in an aqueous solvent.

[0078] In certain embodiments, the composition, when solvated in an aqueous solvent, has a molecular weight of about 3% (w / w) to about 10% (w / w), about 4% (w / w) to about 10% (w / w), about 5% (w / w) to about 10% (w / w), about 6% (w / w) to about 10% (w / w), about 7% (w / w) to about 10% (w / w), about 8% (w / w) to about 10% (w / w), or about 9% (w / w) to about 10% (w / w). 0% (w / w), about 9% (w / w) to about 10% (w / w), about 3% (w / w) to about 9% (w / w), about 3% (w / w) to about 8% (w / w), about 3% (w / w) to about 7% (w / w), about 3% (w / w) to about 6% (w / w), about 3% (w / w) to about 5% (w / w), about 3% (w / w) to about 4% (w / w), about 4% (w / w) to about 9% (w / w), about 4% (w / w) to about 8% (w / w), about 4% (w / w) to about 7% (w / w), about 4% (w / w) to about 6% (w / w), about 4% (w / w) to about 5% (w / w), about 5% (w / w) to about 9% (w / w), about 5% (w / w) to about 8% (w / w), about 5% (w / w) to about 7% (w / w), about The composition may comprise 5% (w / w) to about 6% (w / w), about 6% (w / w) to about 9% (w / w), about 6% (w / w) to about 8% (w / w), about 6% (w / w) to about 7% (w / w), about 7% (w / w) to about 9% (w / w), about 7% (w / w) to about 8% (w / w), or about 8% (w / w) to about 9% (w / w) of maltodextrin. In certain embodiments, the composition comprises about 3% (w / w) to about 10% (w / w) of maltodextrin when solvated in an aqueous solvent.

[0079] In certain embodiments, the composition, when solvated in an aqueous solvent, comprises about 3% (w / w), about 3.5% (w / w), about 4% (w / w), about 4.5% (w / w), about 5% (w / w), about 5.5% (w / w), about 6% (w / w), about 6.5% (w / w), about 7% (w / w), about 7.5% (w / w), about 8% (w / w), about 8.5% (w / w), about 9% (w / w), about 9.5% (w / w), or about 10% (w / w) maltodextrin.

[0080] Oxidation-reduction potential It will be appreciated that cell preservation (e.g., cryopreservation) media have an oxidation-reduction potential that facilitates the viability and growth and / or replication of the microorganisms of interest. The oxidation-reduction potential can be tailored to meet the preferred conditions of a given microorganism or group of microorganisms.

[0081] Anaerobic bacteria (e.g., the microorganisms described herein) require a composition (e.g., a preservation or cryopreservation composition described herein) to be in a reducing state (i.e., a negative redox potential) so that they can be maintained in their natural metabolic state and maintain viability.

[0082] Preparation of the preservation compositions described herein having the desired negative redox potential may be accomplished using one or more of the following techniques: (1) Adding a pre-reducing anaerobic medium to anaerobic microorganisms before or after their growth and / or harvest. The anaerobic medium may contain reducing agents (e.g., cysteine, sodium dithionite, sodium sulfite, thioglycolate, ferric sulfide, dithiothreitol) that can help maintain a reduced environment, as well as redox mediators (e.g., heavy metals (e.g., iron, zinc, manganese, cobalt, copper, nickel, and molybedenum) and vitamins (e.g., menadione, ascorbic acid, folic acid, riboflavin, pyridoxine, biotin, thiamine, nicotinamide, D-pathothenic acid, vitamin b12, lipoic acid, and 4-aminobenzoic acid)). (2) Removal of oxygen from reagents and materials (e.g., growth media and other components) used to make, store, and use the compositions described herein. Such techniques may include, but are not limited to, bubbling an anaerobic inert gas (e.g., nitrogen, carbon dioxide, argon, etc.) through a liquid composition or component blend (see Example 2 disclosed herein). (3) Adding reducing agents (e.g., cysteine, sodium dithionite, sodium sulfite, thioglycolate, ferric sulfide, dithiothreitol) and / or redox mediators (e.g., heavy metals (e.g., iron, zinc, manganese, cobalt, copper, nickel, and molybedenum) and vitamins (e.g., menadione, ascorbic acid, folic acid, riboflavin, pyridoxine, biotin, thiamine, nicotinamide, D-pathothenic acid, vitamin b12, lipoic acid, and 4-aminobenzoic acid)) to the compositions described herein to maintain anaerobicity. (4) Various process steps (e.g., (i) mixing the anaerobic microorganism with the medium, cryopreservative, etc., (ii) pouring the resulting composition into freeze-drying trays, and (iii) freezing the composition) are performed under anaerobic conditions.

[0083] In certain embodiments, the redox potential will be in the range of +300 to -100 mV for facultative anaerobes and +100 mV to about -500 mV for obligate anaerobes. The redox potential is typically measured by a sensing electrode (e.g., a standard hydrogen electrode, a silver chloride electrode, or a saturated calomel electrode. Exemplary redox potential measuring devices include Metler Toledo ORP meters and probes.

[0084] In certain embodiments, the composition may be at any voltage between -50mV and -500mV, -50mV to -450mV, -50mV to -400mV, -50mV to -350mV, -50mV to -300mV, -50mV to -250mV, -50mV to -200mV, -50mV to -150mV, -50mV to -100mV, -100mV to -500mV, -100mV~-450mV, -100mV~-400mV, -100mV~-350mV, -100mV~-300mV, -100mV~-250mV, -100mV~-200mV, -100mV~-150mV, -150mV~-500mV, -150mV~-450mV, -150mV~-400mV, -150mV~-350mV, -150mV~-300mV, - 150mV~-250mV, -150mV~-200mV, -200mV~-500mV, -200mV~-450mV, -200mV~-400mV, -200mV~-350mV, - 200mV~-300mV, -200mV~-250mV, -250mV~-500mV, -250mV~-450mV, -250mV~-400mV, -250mV~-350mV, -2 The composition has a redox potential in the range of 50mV to -300mV, -300mV to -500mV, -300mV to -450mV, -300mV to -400mV, -300mV to -350mV, -350mV to -500mV, -350mV to -450mV, -350mV to -400mV, -400mV to -500mV, -400mV to -450mV, or -450mV to -500mV. In certain embodiments, the composition has a redox potential in the range of -50mV to -500mV when solvated in an aqueous solvent.

[0085] In certain embodiments, the composition may be at about -50mV, about -60mV, about -70mV, about -80mV, about -90mV, about -100mV, about -110mV, about -120mV, about -130mV, about -140mV, about -150mV, about -160mV, about -170mV, about -180mV, for example when solvated in an aqueous solvent. V, about -190 mV, about -200 mV, about -240 mV, about -260 mV, about -280 mV, about -300 mV, about -320 mV, about -340 mV, about -360 mV, about -380 mV, about -400 mV, about -420 mV, about -440 mV, about -460 mV, about -480 mV, or about -500 mV.

[0086] microorganisms In certain embodiments, the composition further comprises one or more anaerobic microorganisms to be preserved. The anaerobic microorganisms can be obligate anaerobes or facultative anaerobes.

[0087] Exemplary anaerobic microorganisms include those from the Bifidobacteriaceae family, Coryobacteriaceae family, Corynebacteriaceae family, Dietziaceae family, Mycobacteriaceae family, Gordoniaceae family, Nocardiaceae family, Streptomycetaceae family, Nocardiaceae family, and the like. 2), Propionibacterineae, Micrococcaceae, Brevibacteriaceae, Dermabacteraceae, Actinomycetaceae, Bogoriellaceae, Promicromonosporaceae, Microbacteriaceae, Cellulomonadaceae, Dermacoccaceae, Intrasporangiaceae, Geodermatophilaceae, Micromonosporaceae, Bacteroidetes, Prevotellaceae, Porphyromonadaceae 1), Rikenellaceae, Porphyromonadaceae 2, Flavobacteriaceae, Sphingobacteriaceae, unclassified Bacteroidetes, Cytophagaceae, Epsilonproteobacteria, Firmicutes, Lactobacillaceae, Leuconostocaceae, Aerococcaceae, Carnobacteriaceae, Aerococcaceae 2, Lachnospiraceae, Streptococci,Enterococcaceae, Erysipelotrichaceae, Staphylococcaceae, XIth family unspecified, Listeriaceae, Planococcaceae 1, XIIth family unspecified, Planococcaceae 2), Bacillaceae, Paenibacillaceae, Clostridiaceae (cluster I), Clostridiaceae (cluster II), Lachnospiraceae (cluster XIVa), Ruminococcaceae (cluster IV), Peptostreptococcaceae (cluster XI), Erysipelothrixaceae (clusters XVII and XVIII), Fusobacteria, Ruminococcaceae, unclassified Clostridiales, Oscillospiraceae, Lachnospiraceae, Eubacteriaceae, family XIII unspecified, Peptostreptococciaceae (cluster XI), Lachnospiraceae (cluster XIVa), Ruminococcaceae (cluster IV), Clostridiaceae (cluster I), Erysipelothrixaceae (cluster XVII and XVIII), I and XVIII), Peptostreptococci, XI Unidentified Family, Erysipelotrichi and Bacillarioles, Clostridia, Negatiwicki, Clostridia 2, Clostridia 3 / Haranaerobiales, Veillonellaceae, Acidaminococcaceae, Erysipelotrichiaceae Cluster XVII, Erysipelotrichiaceae Cluster XVIII ... XVI. Tenericutes, Erysipelothrixaceae, Fusobacteriaceae, Leptotrichaceae, Rhodobacteraceae, Brucellaceae, Rhizobiumaceae, Phyllobacteriaceae, Aurantimonadaceae, Bradyrhizobiaceae, Methylobacteriaceae, Xanthobacteraceae, Caulobacteraceae,Family: Hyphomicrobiaceae, Family: Sphingomonadaceae, Family: Acetobacteraceae, Family: Rhodospirillaceae, Phylum: Firmicutes, Family: Comamonadaceae, Family: Sutterellaceae, Family: Alcaligenaceae, Family: Burkholderiaceae 1, Family: Oxalobacteraceae, Family: Burkholderiaceae 2 ), Rhodocyclaceae, Neisseriaceae, Enterobacteriaceae, Pasteurellaceae, Vibrionaceae, Aeromonadaceae, Succinivibrionaceae, Moraxellaceae, Pseudomonadaceae, Xanthomonadaceae, Sinobacteraceae, ε-Campylobacteraceae 1, ε-Helicobacteraceae, ε-Campylobacteraceae 2), δ-Desulfovibrionaceae, Verrucomicrobia, Lentisphaerae, Planctomycetes, Spirochaetes, Deinococcus-Thermus, Synergistetes, Archaea / Euryarchaeota, Archaea / Crenarchaeota, Fungi-Ascomycota, Fungi-Basidiomycota, Stramenopiles, Apicomplexa, Amoebozoa, Ciliophora,The family includes one or more of the following: Metamonada, Microsporidia, Fornicata, Akkermansiaceae, Christenseneraceae, Odoribacteraceae, Egasellaceae, and combinations thereof.

[0088] In certain embodiments, the one or more anaerobic microorganisms are selected from the group consisting of Akkermansiaceae, Bacillaceae, Bacteroidetaceae, Bifidobacteriaceae, Christenseneraceae, Clostridiaceae, Coriobacteriaceae, Egasellaceae, Enterobacteriaceae, Eubacteriaceae, Lachnospiraceae, Lactobacillaceae, Leuconostocaceae, Odoribacteraceae, Oxalobacteraceae, Porphyromonasaceae, Prevotellaceae, Rikenellaceae, Ruminococcaceae, Streptococciaceae, and combinations thereof.

[0089] The one or more anaerobic microorganisms can be present in the compositions described herein in the form of a cell paste or cell slurry. In certain embodiments, the compositions, when solvated in an aqueous solvent, are present in an amount of about 30% (w / w) to about 50% (w / w), about 32% (w / w) to about 50% (w / w), about 34% (w / w) to about 50% (w / w), about 36% (w / w) to about 50% (w / w), about 38% (w / w) to about 50% (w / w), about 40% (w / w) to about 50% (w / w), about 42% (w / w) to about 50% (w / w), about 44% (w / w) to about 50% (w / w), about 46% (w / w) to about 50% (w / w), about 48% (w / w) to about 50% (w / w). , about 30% (w / w) to about 48% (w / w), about 30% (w / w) to about 46% (w / w), about 30% (w / w) to about 44% (w / w), about 30% (w / w) to about 42% (w / w), about 30% (w / w) to about 40% (w / w), about 30% (w / w) to about 38% (w / w), about 30% (w / w) to about 36% (w / w), about 30% (w / w) to about 34% (w / w), about 30% (w / w) to about 32% (w / w), about 32% (w / w) to about 48% (w / w), about 32% (w / w) to about 46% (w / w), about 32% (w / w) to about 44 %(w / w), about 32%(w / w) to about 42%(w / w), about 32%(w / w) to about 40%(w / w), about 32%(w / w) to about 38%(w / w), about 32%(w / w) to about 36%(w / w), about 32%(w / w) to about 34%(w / w), about 34%(w / w) to about 48%(w / w), about 34%(w / w) to about 46%(w / w), about 34%(w / w) to about 44%(w / w), about 34%(w / w) to about 42%(w / w), about 34%(w / w) to about 40%(w / w), about 34%(w / w) to about 38%(w / w), about 34%(w / w) to about 36% (w / w), about 36% (w / w) to about 48% (w / w), about 36% (w / w) to about 46% (w / w), about 36% (w / w) to about 44% (w / w), about 36% (w / w) to about 42% (w / w), about 36% (w / w) to about 40% (w / w), about 36% (w / w) to about 38% (w / w), about 38% (w / w) to about 48% (w / w), about 38% (w / w) to about 46% (w / w), about 38% (w / w) to about 44% (w / w), about 38% (w / w) to about 42% (w / w), about 38% (w / w) to about 40% (w / w),The cell paste or cell slurry may be about 40% (w / w) to about 48% (w / w), about 40% (w / w) to about 46% (w / w), about 40% (w / w) to about 44% (w / w), about 40% (w / w) to about 42% (w / w), about 42% (w / w) to about 48% (w / w), about 42% (w / w) to about 46% (w / w), about 42% (w / w) to about 44% (w / w), about 44% (w / w) to about 48% (w / w), about 44% (w / w) to about 46% (w / w), or about 46% (w / w) to about 48% (w / w). In certain embodiments, the composition comprises about 30% (w / w) to about 50% (w / w) cell paste or cell slurry when solvated in an aqueous solvent.

[0090] In certain embodiments, the composition, when solvated in an aqueous solvent, comprises about 30% (w / w), about 31% (w / w), about 32% (w / w), about 33% (w / w), about 34% (w / w), about 35% (w / w), about 36% (w / w), about 37% (w / w), about 38% (w / w), about 39% (w / w), about 40% (w / w), about 41% (w / w), about 42% (w / w), about 43% (w / w), about 44% (w / w), about 45% (w / w), about 46% (w / w), about 47% (w / w), about 48% (w / w), about 49% (w / w), or about 50% (w / w) of cell paste or cell slurry.

[0091] In certain embodiments, the one or more anaerobic microorganisms are present in the form of a cell paste, e.g., a compact solid. In certain embodiments, the one or more anaerobic microorganisms are present in the form of a cell slurry, e.g., a cell suspension.

[0092] Stabilizers In certain embodiments, the composition can further comprise a protein stabilizer. In certain embodiments, the composition, when solvated in an aqueous solvent, is from about 5% (w / w) to about 10% (w / w), from about 6% (w / w) to about 10% (w / w), from about 7% (w / w) to about 10% (w / w), from about 8% (w / w) to about 10% (w / w), from about 9% (w / w) to about 10% (w / w), from about 5% (w / w) to about 9% (w / w), from about 5% (w / w) to about 8% (w / w),

[0043] In some embodiments, the composition comprises about 5% (w / w) to about 7% (w / w), about 5% (w / w) to about 6% (w / w), about 6% (w / w) to about 9% (w / w), about 6% (w / w) to about 8% (w / w), about 6% (w / w) to about 7% (w / w), about 7% (w / w) to about 9% (w / w), about 7% (w / w) to about 8% (w / w), or about 8% (w / w) to about 9% (w / w) of the protein stabilizer.

[0093] In certain embodiments, the composition comprises about 5% (w / w), about 5.5% (w / w), about 6% (w / w), about 6.5% (w / w), about 7% (w / w), about 7.5% (w / w), about 8% (w / w), about 8.5% (w / w), about 9% (w / w), about 9.5% (w / w), or about 10% (w / w) of protein stabilizer when solvated in an aqueous solvent.

[0094] In certain embodiments, the protein stabilizing agent is selected from the group consisting of amino acids, antioxidants, disaccharides, reducing agents, polyethylene glycols, and combinations thereof. In certain embodiments, the disaccharides are selected from the group consisting of trehalose, sucrose, isomaltose, lactose, maltose, lactulose, cellobiose, and combinations thereof. In certain embodiments, the protein stabilizing agent comprises trehalose. In certain embodiments, the protein stabilizing agent comprises sucrose and trehalose.

[0095] In certain embodiments, the composition, when solvated in an aqueous solvent, is from about 5% (w / w) to about 10% (w / w), from about 6% (w / w) to about 10% (w / w), from about 7% (w / w) to about 10% (w / w), from about 8% (w / w) to about 10% (w / w), from about 9% (w / w) to about 10% (w / w), from about 5% (w / w) to about 9% (w / w), from about 5% (w / w) to about 8% (w / w), In certain embodiments, the composition comprises from about 5% (w / w) to about 10% (w / w) trehalose when solvated in an aqueous solvent.

[0096] In certain embodiments, the composition, when solvated in an aqueous solvent, comprises about 5% (w / w), about 5.5% (w / w), about 6% (w / w), about 6.5% (w / w), about 7% (w / w), about 7.5% (w / w), about 8% (w / w), about 8.5% (w / w), about 9% (w / w), about 9.5% (w / w), or about 10% (w / w) trehalose.

[0097] In certain embodiments, the composition, when solvated in an aqueous solvent, is from 0% (w / w) to about 10% (w / w), from about 1% (w / w) to about 10% (w / w), from about 2% (w / w) to about 10% (w / w), from about 3% (w / w) to about 10% (w / w), from about 4% (w / w) to about 10% (w / w), from about 5% (w / w) to about 10% (w / w), from about 6% (w / w) to about 10% (w / w), from about 7% (w / w) to about 10% (w / w), from about 8% (w / w) to about 10% (w / w), from about 9% (w / w) to about 10% (w / w), 0% (w / w) to about 9% (w / w), 0% (w / w) ~8%(w / w), 0%(w / w)~7%(w / w), 0%(w / w)~6%(w / w), 0%(w / w)~5%(w / w), 0%(w / w)~about 4%(w / w), 0%(w / w)~about 3%(w / w), 0%(w / w)~about 2%(w / w), 0%(w / w)~about 1% (w / w), about 1% (w / w) to about 9% (w / w), about 1% (w / w) to about 8% (w / w), about 1% (w / w) to about 7% (w / w), about 1% (w / w) to about 6% (w / w), about 1% (w / w) to about 5% (w / w), about 1% (w / w) to about 4% (w / w), about 1% (w / w) to about 3% (w / w), about 1% (w / w) to about 2% (w / w), about 2% (w / w) to about 9% (w / w), about 2% (w / w) to about 8% (w / w), about 2% (w / w) to about 7% (w / w), about 2% (w / w) to about 6% (w / w), about 2% (w / w) to about 5% (w / w), about 2% (w / w) ) to about 4% (w / w), about 2% (w / w) to about 3% (w / w), about 3% (w / w) to about 9% (w / w), about 3% (w / w) to about 8% (w / w), about 3% (w / w) to about 7% (w / w), about 3% (w / w) to about 6% (w / w), about 3% (w / w) to about 5% (w / w), about 3% (w / w) to about 4% (w / w), about 4% (w / w) to about 9% (w / w), about 4% (w / w) to about 8% (w / w), about 4% (w / w) to about 7% (w / w), about 4% (w / w) to about 6% (w / w), about 4% (w / w) to about 5% (w / w), about 5% (w / w) to about 9% (w / w), about 5 %(w / w) to about 8%(w / w), about 5%(w / w) to about 7%(w / w), about 5%(w / w) to about 6%(w / w), about 6%(w / w) to about 9%(w / w), about 6%(w / w) to about 8%(w / w), about 6%(w / w) to about 7%(w / w), about 7%(w / w) to about 9%(w / w),The composition comprises about 7% (w / w) to about 8% (w / w), or about 8% (w / w) to about 9% (w / w) sucrose. In certain embodiments, the composition comprises about 5% (w / w) to about 10% (w / w) sucrose when solvated in an aqueous solvent. In certain embodiments, the composition comprises 0% (w / w) to about 5% (w / w) sucrose when solvated in an aqueous solvent.

[0098] In certain embodiments, the composition, when solvated in an aqueous solvent, comprises about 0.5% (w / w), about 1% (w / w), about 1.5% (w / w), about 2% (w / w), about 2.5% (w / w), about 3% (w / w), about 3.5% (w / w), about 4% (w / w), about 4.5% (w / w), about 5% (w / w), about 5.5% (w / w), about 6% (w / w), about 6.5% (w / w), about 7% (w / w), about 7.5% (w / w), about 8% (w / w), about 8.5% (w / w), about 9% (w / w), about 9.5% (w / w), or about 10% (w / w) sucrose.

[0099] Solvated Compositions In certain embodiments, when the composition is solvated in an aqueous solvent, e.g., water or an aqueous solvent, the composition may be from about 30% (w / w) to about 60% (w / w), from about 35% (w / w) to about 60% (w / w), from about 40% (w / w) to about 60% (w / w), from about 45% (w / w) to about 60% (w / w), from about 50% (w / w) to about 60% (w / w), from about 55% (w / w) to about 60% (w / w), from about 30% (w / w) to about 55% (w / w), from about 30% (w / w) to about 50% (w / w), from about 30% (w / w) to about 45% (w / w), from about 30% (w / w) to about 50% (w / w), The aqueous solvent is about 40% (w / w), about 30% (w / w) to about 35% (w / w), about 35% (w / w) to about 55% (w / w), about 35% (w / w) to about 50% (w / w), about 35% (w / w) to about 45% (w / w), about 35% (w / w) to about 40% (w / w), about 40% (w / w) to about 55% (w / w), about 40% (w / w) to about 50% (w / w), about 40% (w / w) to about 45% (w / w), about 45% (w / w) to about 55% (w / w), about 45% (w / w) to about 50% (w / w), or about 50% (w / w) to about 55% (w / w).

[0100] In certain embodiments, when the composition is solvated in such an aqueous solvent, the composition is about 30% (w / w), about 31% (w / w), about 32% (w / w), about 33% (w / w), about 34% (w / w), about 35% (w / w), about 36% (w / w), about 37% (w / w), about 38% (w / w), about 39% (w / w), about 40% (w / w), about 41% (w / w), about 42% (w / w), about 43% (w / w), about 44% (w / w), about 45% (w / w), about 46% (w / w), about 47% (w / w), about 48% (w / w), about 49% (w / w), about 50% (w / w), about 51% (w / w), about 52% (w / w), about 53% (w / w), about 54% (w / w), about 55% (w / w), about 56% (w / w), about 57% (w / w), about 58% (w / w), about 59% (w / w), about 60% (w / w), about 61% (w / w), about 62% (w / w), about 63% (w / w), about 64% (w / w), about 65% (w / w), about 66% (w / w), about 67% (w / w), about 68% (w / w), about 69% (w / w), about 70% (w / w), about 72% (w / w), about 74% (w / w), about 75% (w / w), about 76% (w / w), about 77% (w / w), about 78% (w / w), about 79% (w / w), about 80% (w / w), about , about 44% (w / w), about 45% (w / w), about 46% (w / w), about 47% (w / w), about 48% (w / w), about 49% (w / w), about 50% (w / w), about 51% (w / w), about 52% (w / w), about 53% (w / w), about 54% (w / w), about 55% (w / w), about 56% (w / w), about 57% (w / w), about 58% (w / w), about 59% (w / w), or about 60% (w / w) of aqueous solvent.

[0101] In certain embodiments, the aqueous solvent is water (e.g., ionized water or deionized water). In certain embodiments, when the composition is solvated in water, the composition may be from about 30% (w / w) to about 60% (w / w), from about 35% (w / w) to about 60% (w / w), from about 40% (w / w) to about 60% (w / w), from about 45% (w / w) to about 60% (w / w), from about 50% (w / w) to about 60% (w / w), from about 55% (w / w) to about 60% (w / w), from about 30% (w / w) to about 55% (w / w), from about 30% (w / w) to about 50% (w / w), from about 30% (w / w) to about 45% (w / w), from about 30% (w / w) to about 40% (w / w).

[0043] In one embodiment, the composition comprises about 30% (w / w) to about 35% (w / w), about 35% (w / w) to about 55% (w / w), about 35% (w / w) to about 50% (w / w), about 35% (w / w) to about 45% (w / w), about 35% (w / w) to about 40% (w / w), about 40% (w / w) to about 55% (w / w), about 40% (w / w) to about 50% (w / w), about 40% (w / w) to about 45% (w / w), about 45% (w / w) to about 55% (w / w), about 45% (w / w) to about 50% (w / w), or about 50% (w / w) to about 55% (w / w) of water.

[0102] In certain embodiments, when the composition is solvated in water, the composition is about 30% (w / w), about 31% (w / w), about 32% (w / w), about 33% (w / w), about 34% (w / w), about 35% (w / w), about 36% (w / w), about 37% (w / w), about 38% (w / w), about 39% (w / w), about 40% (w / w), about 41% (w / w), about 42% (w / w), about 43% (w / w), about 44% (w / w), about 45% (w / w), about 46% (w / w), about 47% (w / w), about 48% (w / w), about 49% (w / w), about 50% (w / w), about 51% (w / w), about 52% (w / w), about 53% (w / w), about 54% (w / w), about 55% (w / w), about 56% (w / w), about 57% (w / w), about 58% (w / w), about 59% (w / w), or about 60% (w / w) water.

[0103] Exemplary Preservative Compositions In various embodiments, provided herein is a composition for preserving anaerobic microorganisms that includes, inter alia, a viscosity modifier, an energy source for the microorganisms, and an aqueous solvent.

[0104] In various embodiments, the composition for preserving anaerobic microorganisms comprises: (i) about 3% (w / w) to about 15% (w / w) of an energy source for the microorganisms; (ii) about 0.01% (w / w) to about 0.4% (w / w) of a viscosity modifier; and (iii) about 30% (w / w) to about 60% (w / w) of an aqueous solvent; Includes.

[0105] In various embodiments, the composition comprises a viscosity modifier, an energy source for the microorganism, and an aqueous solvent, and has a viscosity in the range of about 5 cP to about 6,000 cP, an osmolality in the range of about 300 Osm / L to about 4,000 Osm / L, a pH in the range of about 4 to about 8, and a redox potential in the range of about -50 mV to about -500 mV.

[0106] In various embodiments, the composition for preserving anaerobic microorganisms comprises: (i) about 3% (w / w) to about 15% (w / w) of an energy source for the microorganisms; (ii) about 0.01% (w / w) to about 0.4% (w / w) of a viscosity modifier; and (iii) about 30% (w / w) to about 60% (w / w) of an aqueous solvent; and has a viscosity in the range of about 5 cP to about 6,000 cP, an osmolality in the range of about 300 Osm / L to about 4,000 Osm / L, a pH in the range of about 4 to about 8, and a redox potential in the range of about -50 mV to about -500 mV.

[0107] In various embodiments, the composition comprises: (i) a cell paste containing about 30% (w / w) to about 50% (w / w) of one or more anaerobic microorganisms; (ii) about 3% (w / w) to about 15% (w / w) of an energy source for the microorganisms; (iii) about 0.01% (w / w) to about 0.4% (w / w) of a viscosity modifier; and (iv) about 30% (w / w) to about 60% (w / w) of an aqueous solvent; Includes.

[0108] In various embodiments, the composition comprises: (i) a cell paste containing about 30% (w / w) to about 50% (w / w) of one or more anaerobic microorganisms; (ii) about 3% (w / w) to about 15% (w / w) of an energy source for the microorganisms; (iii) about 0.01% (w / w) to about 0.4% (w / w) of a viscosity modifier; and (iv) about 30% (w / w) to about 60% (w / w) of an aqueous solvent; and has a viscosity in the range of about 5 cP to about 6,000 cP, an osmolality in the range of about 300 Osm / L to about 4,000 Osm / L, a pH in the range of about 4 to about 8, and a redox potential in the range of about -50 mV to about -500 mV.

[0109] In various embodiments, the composition comprises: (i) a cell paste containing about 30% (w / w) to about 50% (w / w) of one or more anaerobic microorganisms; (ii) about 0.5% (w / w) to about 5% (w / w) of raftilose; (iii) about 3% (w / w) to about 10% (w / w) of maltodextrin; (iv) about 0% (w / w) to about 0.4% (w / w) sodium alginate; (v) about 5% (w / w) to about 10% (w / w) trehalose; (vi) 0% (w / w) to about 5% (w / w) sucrose; (vii) about 35% (w / w) to about 55% (w / w) water; Includes.

[0110] In various embodiments, the composition comprises: (i) a cell paste containing about 30% (w / w) to about 50% (w / w) of one or more anaerobic microorganisms; (ii) about 0.5% (w / w) to about 5% (w / w) of raftilose; (iii) about 3% (w / w) to about 10% (w / w) of maltodextrin; (iv) about 0% (w / w) to about 0.4% (w / w) sodium alginate; (v) about 5% (w / w) to about 10% (w / w) trehalose; (vi) 0% (w / w) to about 5% (w / w) sucrose; (vii) about 35% (w / w) to about 55% (w / w) water; and has a viscosity in the range of about 5 cP to about 6,000 cP, an osmolality in the range of about 300 Osm / L to about 4,000 Osm / L, a pH in the range of about 4 to about 8, and a redox potential in the range of about -50 mV to about -500 mV.

[0111] In various embodiments, provided herein is a cryopreservation composition for cryopreserving anaerobic microorganisms, the cryopreservation composition comprising a viscosity modifier, an energy source for the microorganism, and an aqueous solvent, the cryopreservation composition having a viscosity in the range of about 5 cP to about 6,000 cP, an osmolality in the range of about 300 Osm / L to about 4,000 Osm / L, a pH in the range of about 4 to about 8, and an oxidation-reduction potential in the range of about -50 mV to about -500 mV.

[0112] In various embodiments, (i) about 3% (w / w) to about 15% (w / w) of an energy source for the microorganisms; (ii) about 0.01% (w / w) to about 0.4% (w / w) of a viscosity modifier; and (iii) 30% (w / w) to about 60% (w / w) of an aqueous solvent; Provided herein is a cryopreservation composition for cryopreserving anaerobic microorganisms, comprising:

[0113] In various embodiments, (i) about 3% (w / w) to about 15% (w / w) of an energy source for the microorganisms; (ii) about 0.01% (w / w) to about 0.4% (w / w) of a viscosity modifier; and (iii) 30% (w / w) to about 60% (w / w) of an aqueous solvent; Provided herein is a cryopreservation composition for cryopreserving anaerobic microorganisms, comprising:

[0114] In various embodiments, (i) a cell paste containing about 30% (w / w) to about 50% (w / w) of one or more anaerobic microorganisms; (ii) about 3% (w / w) to about 15% (w / w) of an energy source for the microorganisms; (iii) about 0.01% (w / w) to about 0.4% (w / w) of a viscosity modifier; and (iv) about 30% (w / w) to about 60% (w / w) of an aqueous solvent; Provided herein is a cryopreservation composition for cryopreserving anaerobic microorganisms, comprising:

[0115] In various embodiments, (i) a cell paste containing about 30% (w / w) to about 50% (w / w) of one or more anaerobic microorganisms; (ii) about 3% (w / w) to about 15% (w / w) of an energy source for the microorganisms; (iii) about 0.01% (w / w) to about 0.4% (w / w) of a viscosity modifier; and (iv) about 30% (w / w) to about 60% (w / w) of an aqueous solvent; Provided herein is a cryopreservation composition for cryopreserving anaerobic microorganisms, comprising:

[0116] In various embodiments, (i) a cell paste containing about 30% (w / w) to about 50% (w / w) of one or more anaerobic microorganisms; (ii) about 0.5% (w / w) to about 5% (w / w) of raftilose; (iii) about 3% (w / w) to about 10% (w / w) of maltodextrin; (iv) about 0% (w / w) to about 0.4% (w / w) sodium alginate; (v) about 5% (w / w) to about 10% (w / w) trehalose; (vi) 0% (w / w) to about 5% (w / w) sucrose; (vii) about 35% (w / w) to about 55% (w / w) water; Provided herein is a cryopreservation composition for cryopreserving anaerobic microorganisms, comprising:

[0117] In various embodiments, (i) a cell paste containing about 30% (w / w) to about 50% (w / w) of one or more anaerobic microorganisms; (ii) about 0.5% (w / w) to about 5% (w / w) of raftilose; (iii) about 3% (w / w) to about 10% (w / w) of maltodextrin; (iv) about 0% (w / w) to about 0.4% (w / w) sodium alginate; (v) about 5% (w / w) to about 10% (w / w) trehalose; (vi) 0% (w / w) to about 5% (w / w) sucrose; (vii) about 35% (w / w) to about 55% (w / w) water; Provided herein is a cryopreservation composition for cryopreserving anaerobic microorganisms, comprising:

[0118] In certain embodiments, the preservation compositions (e.g., cryopreservation compositions) described herein are not cross-linked compositions or do not contain cross-linked components, hi certain embodiments, the compositions are not glass or glass-like.

[0119] 2. Preservation composition In one aspect, provided herein is a preservation composition that generally comprises one or more anaerobic microorganisms, a viscosity modifier, and an energy source for the microorganism. In certain embodiments, the preservation composition has a moisture content of less than 5% (w / w).

[0120] In various embodiments, provided herein are preservation compositions, e.g., cryopreservation compositions, comprising one or more anaerobic microorganisms, a viscosity modifier, an energy source for the microorganisms, and a moisture content of less than 5% (w / w).

[0121] In certain embodiments, the preservation composition is a cryopreservation composition.

[0122] In certain embodiments, the one or more anaerobic microorganisms can be facultative anaerobes, obligate anaerobes, or a combination thereof. Anaerobic microorganisms include Bifidobacteriaceae, Coryobacteriaceae, Corynebacteriaceae, Diesiaceae, Mycobacteriaceae, Gordoniaceae, Nocardiaceae, Streptomycetaceae, Nocardiaceae, Propionibacteriaceae, Micrococcaceae, Brevibacteriaceae, Dermabacteraceae, Actinomycetaceae, Bogoriellaceae, Promicromonosporaceae, Microbacteriaceae, Cellulomonasaceae, Dermacocciaceae, Intrasporangiaceae, Geodermatraphiaceae, Micromonosporaceae, Bacteroides, and the like. Iodesaceae, Prevotellaceae, Porphyromonas 1, Rikenellaceae, Porphyromonas 2, Flavobacteriaceae, Sphingobacteriaceae, unclassified Bacteroidetes, Cytophagaceae, Epsilonproteobacteria, Firmicutes, Lactobacillaceae, Leuconostocaceae, Aerococcaceae, Carnobacteriaceae, Aerococcaceae 2, Lachnospiraceae, Streptococciaceae, Enterococciaceae, Erysipelothrixaceae, Staphylococciaceae, XIth family unspecified, Listeriaceae, Planococcus 1, Families, XII unknown families, Planococciaceae, Bacillaceae, Paniebacillaceae, Clostridiaceae (cluster I), Clostridiaceae (cluster II), Lachnospiraceae (cluster XIVa), Ruminococcaceae (cluster IV), Peptostreptococciaceae (cluster XI), Erysipelothrixaceae (clusters XVII and XVIII), Fusobacteriaceae, Ruminococcaceae, unclassified Clostridiales, Oscillospiraceae, Lachnospiraceae, Eubacteriaceae, XIII unknown families Acrocephalidae, Peptostreptococci (cluster XI), Lachnospiraceae (cluster XIVa), Ruminococcaceae (cluster IV), Clostridia (cluster I), Erysipelotrichaceae (clusters XVII and XVIII), Peptostreptococci, XI Unidentified Family, Erysipelotrichidae and Bacilli, Clostridia, Negativicutes, Clostridia 2 (Cluster XI),2), Clostridia 3 / Halanaerobiales, Veillonellaceae, Acidaminococcaceae, Erysipelotrichidae Cluster XVII, Erysipelotrichidae Cluster XVIII, Erysipelotrichidae Cluster XVI, Tenericutes, Erysipelotrichidae, Fusobacteriaceae, Leptotrichidae Leptotrichiaceae, Rhodobacteraceae, Brucellaceae, Rhizobiaceae, Phyllobacteriaceae, Aurantimonadaceae, Brassirhizobiumaceae, Methylobacteraceae, Xanthobacteraceae, Caulobacteraceae, Hyphomicrobiumaceae, Sphingomonasaceae, Acetobacteriaceae, Rhodobacteraceae, Spirillaceae, Firmicutes, Comamonadaceae, Saterellaceae, Alcaligenesaceae, Burkholideriaceae, Oxalobacteraceae, Burkholideriaceae II, Rhodocyclaceae, Neisseriaceae, Enterobacteriaceae, Pasteurellaceae, Vibriaceae, Aeromonadaceae, Succinivibrionaceae, Moraxellaceae, Pseudomonadaceae, Xanthomonadaceae, Sinobacteraceae, ε-Campylobacteridae, ε-Helicobacteraceae, ε-Campylobacteridae II, δ-Desulfovibrioaceae, Vermicobium , Lentispaella, Planctomycetes, Spirochaetae, Deinococcus-Thermus, Synergistea, Archaea / Euryarchaeota, Archaea / Crenarchaeota, Fungi-Ascomycota, Fungi-Basidiomycota, Stramenopiles, Apicomplexa, Amoebozoa, Ciliata, Metamonadena, Microsporidia, Fornicata, Akkermansia, Christenseneraceae, Odoribacteraceae, Egasellaceae, and combinations thereof.

[0123] In certain embodiments, the one or more anaerobic microorganisms may be selected from the group consisting of Akkermansiaceae, Bacillaceae, Bacteroidetaceae, Bifidobacteriaceae, Christensellaceae, Clostridiaceae, Coriobacteriaceae, Egasellaceae, Enterobacteriaceae, Eubacteriaceae, Lachnospiraceae, Lactobacillaceae, Leuconostocaceae, Odoribacteraceae, Oxalobacteraceae, Porphyromonasaceae, Prevotellaceae, Rikenellaceae, Ruminococcaceae, Streptococciaceae, and combinations thereof.

[0124] In certain embodiments, the composition has a water content of less than about 5% (w / w), less than about 4.5% (w / w), less than about 4% (w / w), less than about 3.5% (w / w), less than about 3% (w / w), less than about 2.5% (w / w), less than about 2% (w / w), less than about 1.5% (w / w), less than about 1% (w / w), or less than about 0.5% (w / w).

[0125] In certain embodiments, the composition is from about 1% (w / w) to about 5% (w / w), from about 1.5% (w / w) to about 5% (w / w), from about 2% (w / w) to about 5% (w / w), from about 2.5% (w / w) to about 5% (w / w), from about 3% (w / w) to about 5% (w / w), from about 3.5% (w / w) to about 5% (w / w), from about 4% (w / w) to about 5% (w / w), from about 4.5% (w / w) to about 5% (w / w), from about 1% (w / w) to about 4.5% (w / w) / w), about 1% (w / w) to about 4% (w / w), about 1% (w / w) to about 3.5% (w / w), about 1% (w / w) to about 3% (w / w), about 1% (w / w) to about 2.5% (w / w), about 1% (w / w) to about 2% (w / w), about 1% (w / w) to about 1.5% (w / w), about 1.5% (w / w) to about 4.5% (w / w), about 1.5% (w / w) to about 4% (w / w), about 1.5% (w / w) to about 3.5% (w / w), about 1.5 %(w / w) to about 3%(w / w), about 1.5%(w / w) to about 2.5%(w / w), about 1.5%(w / w) to about 2%(w / w), about 2%(w / w) to about 4.5%(w / w), about 2%(w / w) to about 4%(w / w), about 2%(w / w) to about 3.5%(w / w), about 2%(w / w) to about 3%(w / w), about 2%(w / w) to about 2.5%(w / w), about 2.5%(w / w) to about 4.5%(w / w), about 2.5%(w / w) The composition has a water content in the range of about 4% (w / w), about 2.5% (w / w) to about 3.5% (w / w), about 2.5% (w / w) to about 3% (w / w), about 3% (w / w) to about 4.5% (w / w), about 3% (w / w) to about 4% (w / w), about 3% (w / w) to about 3.5% (w / w), about 3.5% (w / w) to about 4.5% (w / w), about 3.5% (w / w) to about 4% (w / w), or about 4% (w / w) to about 4.5% (w / w). In certain embodiments, the composition has a water content in the range of about 2% (w / w) to about 4% (w / w) or about 2% (w / w) to about 3% (w / w). In certain embodiments, the composition has a moisture content in the range of about 1.5% (w / w) to about 4% (w / w).

[0126] In certain embodiments, the composition has a moisture content of about 0.5% (w / w), about 1% (w / w), about 1.5% (w / w), about 2% (w / w), about 2.5% (w / w), about 3% (w / w), about 3.5% (w / w), about 4% (w / w), about 4.5% (w / w), or about 5% (w / w).

[0127] In certain embodiments, the viscosity modifier is a polymer composition, which may include alginate, acacia, alginic acid, agar, aramic acid, bentonite, carbomer, carrageenan, cellulose, dextrin, gelatin, gellan gum, guar gum, carboxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, methylcellulose, mucin, pectin, starch, xanthan gum, or a combination thereof.

[0128] In certain embodiments, the polymer composition includes an alginate, such as sodium alginate. In certain embodiments, sodium alginate has a viscosity of about 250 cP to about 14,000 cP, about 500 cP to about 14,000 cP, about 750 cP to about 14,000 cP, about 1,000 cP to about 14,000 cP, about 2,000 cP to about 14,000 cP, about 3,000 cP to about 14,000 cP, about 4,000 cP to about 14,000 cP, about 5,000 cP to about 14,000 cP, about 6,000 cP to about 14,000 cP, about 7,000 cP to about 14,000 cP, about 8,000 cP to about 14,000 cP, about 9,000 cP to about 14,000cP, approx. 10,000cP ~ approx. 14,000cP, approx. 11,000cP ~ approx. 14,000cP, approx. 12,000cP ~ Approx. 14,000cP, Approx. 13,000cP~Approx. 14,000cP, Approx. 250cP~Approx. 13,000cP, Approx. 250cP~Approx. 12,00 0cP, about 250cP to about 11,000cP, about 250cP to about 10,000cP, about 250cP to about 9,000cP, about 250c P ~ about 8,000cP, about 250cP - about 7,000cP, about 250cP - about 6,000cP, about 250cP - about 5,000cP, about 250cP~Approx. 4,000cP, Approx. 250cP~Approx. 3,000cP, Approx. 250cP~Approx. 2,000cP, Approx. 250cP~Approx. 1,00 0cP, about 250cP to about 750cP, about 250cP to about 500cP, about 500cP to about 13,000cP, about 500cP to about 12 ,000cP, about 500cP to about 11,000cP, about 500cP to about 10,000cP, about 500cP to about 9,000cP, about 5 00cP ~ approx. 8,000cP, approx. 500cP ~ approx. 7,000cP, approx. 500cP ~ approx. 6,000cP, approx. 500cP ~ approx. 5,000c P, about 500cP to about 4,000cP, about 500cP to about 3,000cP, about 500cP to about 2,000cP, about 500cP to about 1 ,000cP, about 500cP to about 750cP, about 750cP to about 13,000cP, about 750cP to about 12,000cP, about 750c P ~ approx. 11,000cP, approx. 750cP ~ approx. 10,000cP, approx. 750cP ~ approx. 9,000cP, approx. 750cP ~ approx. 8,000c P, about 750cP to about 7,000cP, about 750cP to about 6,000cP, about 750cP to about 5,000cP, about 750cP to about 4,000 cP, from about 750 cP to about 3,000 cP, from about 750 cP to about 2,000 cP, from about 750 cP to about 1,000 cP, from about 1,000 cP to about 13,000 cP, from about 1,000 cP to about 12,000 cP, from about 1,000 cP to about 11,000 cP, from about 1,000 cP to about 10,000 cP, from about 1,000 cP to about 9,000 cP, from about 1,000 cP to about 8,000 cP, from about 1,000 cP to about 7,000 cP, from about 1,000 cP to about 6,000 cP, from about 1,000 cP to about 5,000 cP, from about 1,000 cP to about 4,000 cP, from about 1,000 cP to about 3,000 cP, from about 1,000 cP to about 2,000 cP, from about 2,000 cP to about 13,000 cP, from about 2,000 cP to about 12,000 cP, from about 2,000 cP to about 11,000 cP, from about 2,000 cP to about 10,000 cP, from about 2,000 cP to about 9,000 cP, from about 2,000 cP to about 8,000 cP, from about 2,000 cP to about 7,000 cP, from about 2,000 cP to about 6,000 cP, from about 2,000 cP to about 5,000 cP, from about 2,000 cP to about 4,000 cP, from about 2,000 cP to about 3,000 cP, from about 3,000 cP to about 13,000 cP, from about 3,000 cP to about 12,000 cP, from about 3,000 cP to about 11,000 cP, from about 3,000 cP to about 10,000 cP, from about 3,000 cP to about 9,000 cP, from about 3,000 cP to about 8,000 cP, from about 3,000 cP to about 7,000 cP, from about 3,000 cP to about 6,000 cP, from about 3,000 cP to about 5,000 cP, from about 3,000 cP to about 4,000 cP, from about 4,000 cP to about 13,000 cP, from about 4,000 cP to about 12,000 cP, from about 4,000 cP to about 11,000 cP, from about 4,000 cP to about 10,000 cP, from about 4,000 cP to about 9,000 cP, from about 4,000 cP to about 8,000 cP, from about 4,000 cP to about 7,000 cP, from about 4,000 cP to about 6,000 cP, from about 4,000 cP to about 5,000 cP, from about 5,000 cP to about 13,000 cP, from about 5,000 cP to about 12,000 cP, from about 5,000 cP to about 11,000 cP, from about 5,000 cP to about 10,000 cP, from about 5,000 cP to about 9,000 cP, from about 5,000 cP to about 8,000 cP, from about 5,000 cP to about 7,000 cP, from about 5,000 cP to about 6,000 cP, from about 6,000 cP to about 13,000 cP, from about 6,000cP ~ approx. 12,000cP, approx. 6,000cP ~ approx. 11,000cP, approx. 6,000cP ~ approx. 10,000cP, approx. 6,000cP ~ approx. 9 ,000cP, about 6,000cP to about 8,000cP, about 6,000cP to about 7,000cP, about 7,000cP to about 13,000cP, about 7, 000cP ~ approx. 12,000cP, approx. 7,000cP ~ approx. 11,000cP, approx. 7,000cP ~ approx. 10,000cP, approx. 7,000cP ~ approx. 9 ,000cP, approx. 7,000cP ~ approx. 8,000cP, approx. 8,000cP ~ approx. 13,000cP, approx. 8,000cP ~ approx. 12,000cP, approx. 8, 000cP ~ approx. 11,000cP, approx. 8,000cP ~ approx. 10,000cP, approx. 8,000cP ~ approx. 9,000cP, approx. 9,000cP ~ approx. 13 ,000cP, about 9,000cP to about 12,000cP, about 9,000cP to about 11,000cP, about 9,000cP to about 10,000cP, about 1 The viscosity ranges from about 0,000 cP to about 13,000 cP, about 10,000 cP to about 12,000 cP, about 10,000 cP to about 11,000 cP, about 11,000 cP to about 13,000 cP, about 11,000 cP to about 12,000 cP, or about 12,000 cP to about 13,000 cP. In certain embodiments, sodium alginate has a viscosity range of about 250 cP to about 14,000 cP.

[0129] In certain embodiments, the energy source is a cell culture medium that allows the growth of the microorganism. In certain embodiments, the energy source is a monosaccharide, a disaccharide, an oligosaccharide, or a combination thereof. In certain embodiments, the monosaccharide is selected from the group consisting of glucose, fructose, xylose, and combinations thereof. In certain embodiments, the disaccharide is trehalose, sucrose, or a combination thereof. In certain embodiments, the oligosaccharide is an inulin derivative. In certain embodiments, the inulin derivative is a fructooligosaccharide. In certain embodiments, the fructooligosaccharide is raftilose. In certain embodiments, the oligosaccharide is maltodextrin.

[0130] In certain embodiments, the energy source comprises raftilose and maltodextrin.

[0131] In certain embodiments, the composition further comprises a protein stabilizing agent, hi certain embodiments, the protein stabilizing agent is selected from the group consisting of amino acids, antioxidants, disaccharides, reducing agents, polyethylene glycol, and combinations thereof.

[0132] In certain embodiments, the disaccharide is selected from the group consisting of trehalose, sucrose, isomaltose, lactose, maltose, lactulose, cellobiose, and combinations thereof. In certain embodiments, the protein stabilizing agent comprises trehalose. In certain embodiments, the protein stabilizing agent comprises sucrose and trehalose.

[0133] In certain embodiments, the composition further comprises a buffering agent.

[0134] In certain embodiments, the preservation composition comprises: (i) one or more anaerobic microorganisms; (ii) an energy source comprising raftilose and maltodextrin; (iii) sodium alginate; (iv) protein stabilizing agents including trehalose, sucrose, and combinations thereof; Includes.

[0135] In certain embodiments, the preservation composition comprises: (i) one or more anaerobic microorganisms; (ii) an energy source comprising raftilose and maltodextrin; (iii) sodium alginate; (iv) a protein stabilizing agent comprising trehalose, sucrose, and combinations thereof; (v) a water content of less than 5% (w / w); Includes.

[0136] In certain embodiments, the preservation composition comprises: (i) one or more anaerobic microorganisms; (ii) an energy source comprising raftilose and maltodextrin; (iii) sodium alginate; (iv) protein stabilizing agents including trehalose, sucrose, and combinations thereof; Includes.

[0137] In certain embodiments, the preservation composition comprises: (i) one or more anaerobic microorganisms; (ii) an energy source comprising raftilose and maltodextrin; (iii) sodium alginate; (iv) a protein stabilizing agent comprising trehalose, sucrose, and combinations thereof; (v) a water content of less than 5% (w / w); Includes.

[0138] In certain embodiments, the composition, when stored at 4° C. for 6 or 12 months and then rehydrated, results in less than 3, 2, 1, or 0.5 log loss of colony forming units (CFU). In certain embodiments, the composition, when stored at 4° C. for 6 or 12 months and then rehydrated, results in less than 3 log loss of colony forming units (CFU). In certain embodiments, the composition, when stored at 4° C. for 6 or 12 months and then rehydrated, results in less than 2 log loss of colony forming units (CFU). In certain embodiments, the composition, when stored at 4° C. for 6 or 12 months and then rehydrated, results in less than 1 log loss of colony forming units (CFU). In certain embodiments, the composition, when stored at 4° C. for 6 or 12 months and then rehydrated, results in less than 0.5 log loss of colony forming units (CFU).

[0139] In certain embodiments, the composition remains substantially stable (e.g., less than 0.5 log loss in potency (CFU / g)) for about 0.5 years, about 1 year, about 1.5 years, about 2 years, about 2.5 years, or about 3 years at 4° C. In certain embodiments, the composition remains substantially stable for about 2 years at 4° C.

[0140] In certain embodiments, the composition, when rehydrated in an aqueous fluid, allows for the regrowth of one or more microorganisms within 2, 3, 4, or 5 hours of rehydration. In certain embodiments, the composition, when rehydrated in an aqueous fluid, allows for the regrowth of one or more microorganisms within 2 hours of rehydration. In certain embodiments, the composition, when rehydrated in an aqueous fluid, allows for the regrowth of one or more microorganisms within 3 hours of rehydration. In certain embodiments, the composition, when rehydrated in an aqueous fluid, allows for the regrowth of one or more microorganisms within 4 hours of rehydration. In certain embodiments, the composition, when rehydrated in an aqueous fluid, allows for the regrowth of one or more microorganisms within 5 hours of rehydration.

[0141] In certain embodiments, the preservation compositions (e.g., cryopreservation compositions) described herein are not cross-linked compositions, hi certain embodiments, the compositions are not glass or glass-like.

[0142] 3. Pharmaceutical compositions and food products In one aspect, provided herein are pharmaceutical compositions generally comprising a preservation composition or a cryopreservation composition described herein, and optionally comprising one or more pharma- ceutically acceptable excipients.

[0143] In various embodiments, provided herein is a pharmaceutical composition comprising a preservation composition (e.g., a cryopreservation composition) described herein and one or more pharma- ceutically acceptable excipients.

[0144] The pharmaceutical compositions provided herein can be administered by various routes, including but not limited to oral (enteral), parenteral (by injection), or rectal administration.In certain embodiments, the pharmaceutical compositions disclosed herein are administered orally.In certain embodiments, the pharmaceutical compositions described herein are administered by injection, for example, via subcutaneous or intramuscular injection, or by injection into tissue requiring medical treatment, for example, cancerous tissue.

[0145] The pharmaceutical compositions provided herein may also be administered chronically ("chronic administration"). Chronic administration refers to administration of a compound or pharmaceutical composition thereof that can continue for an extended period of time, for example, 3 months, 6 months, 1 year, 2 years, 3 years, 5 years, etc., or indefinitely, for example, for the rest of the subject's life. In certain embodiments, chronic administration is intended to provide a constant level of one or more microorganisms described herein in the gastrointestinal tract of a subject.

[0146] The pharmaceutical composition provided herein can be presented in unit dosage form to facilitate accurate dosing.The term "unit dosage form" refers to a physically separate unit suitable for human subjects and other mammals as a unit dosage, each unit containing a predetermined amount of active material calculated to produce desired therapeutic effect in association with suitable pharmaceutical excipients.Typical unit dosage forms include pre-filled, pre-measured ampoules or syringes of liquid compositions, or pills, tablets, capsules, etc., for solid compositions.

[0147] In certain embodiments, the pharmaceutical compositions provided herein are administered to patients as solid dosage forms. In certain embodiments, the solid dosage form is a tablet. In certain embodiments, the solid dosage form is a capsule. In certain embodiments, the solid dosage form is a caplet.

[0148] Although the description of pharmaceutical compositions provided herein is directed primarily to pharmaceutical compositions suitable for administration to humans, those skilled in the art will understand that such compositions are generally suitable for administration to any animal. Modifications of pharmaceutical compositions suitable for administration to humans to make the compositions suitable for administration to various animals are well understood, and a veterinary pharmacologist of ordinary skill can design and / or make such modifications using routine experimentation. General considerations in the formulation and / or manufacture of pharmaceutical compositions can be found, for example, in REMINGTON: THE SCIENCE AND PRACTICE OF PHARMACE, 21st Edition, Lippincott Williams&Wilkins, 2005. In certain embodiments, the pharmaceutical composition is placed in a capsule, tablet, or caplet.

[0149] In another aspect, provided herein are food products and probiotics, generally comprising the preserved or cryopreserved compositions described herein. In certain embodiments, the food product or probiotics are disposed within a capsule, tablet, or caplet.

[0150] In certain circumstances, it may be desirable to administer the pharmaceutical composition or food product / probiotic by oral delivery. To deliver the microorganisms to the intestinal tract having a suitable environment (e.g., redox potential) for the recovery and growth of the microorganisms, it may be desirable to coat the capsule, tablet, or caplet with a functional coating, e.g., a coating that releases the microorganisms in a suitable environment (e.g., an environment having a suitable pH and / or redox potential) to promote the recovery and growth of the microorganisms.

[0151] For example, capsules, tablets, or caplets may be coated with a delayed release coating, such as an enteric coating. Enteric coatings are generally acid-resistant, which protect the components of the dosage form from release in the stomach, yet dissolve or disintegrate under the neutral or slightly alkaline conditions typically encountered after the stomach. Exemplary coatings include hydroxypropylmethylcellulose phthalate, methacrylic acid / methumethyl methacrylate polymers (e.g., Eudragit® materials such as Eudragit® L100-55, Eudragit® L30D, and Eudragit® S-100), cellulose acetate phthalate, cellulose acetate, trimellitate, hydroxypropylmethylcellulose phthalate, hydroxypropylmethylcellulose acetate succinate, poly(MA-EA) * 1:1, Poly(MA-MMA) * 1:1, Poly(MA-MMA) * 1:2, HP-F, sureteric, acryl-Eze, Advantia performance, aquateric, aquocoat ECD, aqoat, CAP, CAT, and polyvinyl acetate phthalate (see, e.g., U.S. Pat. Nos. 9,629,803 B2 and US2018 / 0325825). These coated capsules, tablets, and caplets allow their contents to remain dry until the capsules, tablets, and caplets reach a suitable environment, e.g., pH and / or redox potential, where the coating degrades and releases the microorganisms into the desired environment in the subject to promote recovery and growth.

[0152] Additionally, optionally, the cryopreserved organisms can be mixed with a carbonated solution prior to ingestion. When ingested orally without food (e.g., on an empty stomach), the organisms have a short residence time in the stomach and can enter the intestinal tract within 30 minutes of ingestion.

[0153] 4. Method for preparing cryopreservation compositions of anaerobic microorganisms The present disclosure also provides methods of producing the cryopreserved compositions described herein.

[0154] In certain embodiments, the method generally comprises two steps: the first step is combining an anaerobic microorganism with an excipient blend comprising an aqueous mixture comprising a viscosity modifier and an energy source for the microorganism to produce a cryopreservation mixture, the cryopreservation mixture having a viscosity in the range of about 5 cP to about 6,000 cP, an osmolality in the range of about 300 Osm / L to about 4,000 Osm / L, a pH in the range of about 4 to about 8, and optionally a redox potential in the range of -50 mV to about -500 mV. Then, in a separate step, the cryopreservation mixture is lyophilized to reduce the water content of the cryopreservation mixture to less than about 5% (w / w) to produce a cryopreservation composition.

[0155] In certain embodiments, during the first step (step (a)), the anaerobic microorganism is selected from the group consisting of Bifidobacteriaceae, Coriobacteriaceae, Corynebacteriaceae, Diesiaceae, Mycobacteriaceae, Gordoniaceae, Nocardiaceae, Streptomycetaceae, Nocardiaceae, Propionibacteriaceae, Micrococcaceae, Brevibacteriaceae, Dermabacteraceae, Actinomycetaceae, Bogoriellaceae, Promicromonosporaceae, Microbacteriaceae, Cellulomonasaceae, Dermacococciaceae, Intrasporangiaceae, Geodermatraphis, Families: Micromonosporaceae, Bacteroidetes, Prevotellaceae, Porphyromonas 1, Rikenellaceae, Porphyromonas 2, Flavobacteriaceae, Sphingobacteriaceae, Unclassified Bacteroidetes, Cytophagaceae, Class Epsilonproteobacteria, Phylum Firmicutes, Lactobacillaceae, Leuconostocaceae, Aerococcaceae, Carnobacteriaceae, Aerococcaceae 2, Lachnospiraceae, Streptococciaceae, Enterococciaceae, Erysipelothrixaceae, Staphylococciaceae, Family XI Unspecified, Listeriaceae, Family Planococci 1 , XII unknown family, Planococciaceae, Bacillaceae, Paniebacillaceae, Clostridiaceae (cluster I), Clostridiaceae (cluster II), Lachnospiraceae (cluster XIVa), Ruminococcaceae (cluster IV), Peptostreptococciaceae (cluster XI), Erysipelothrixaceae (clusters XVII and XVIII), Fusobacteriaceae, Ruminococcaceae, unclassified Clostridiales, Oscillospiraceae, Lachnospiraceae, Eubacteriaceae, XIII unknown family, Peptostreptococciaceae (Cluster XVII and XVIII), star XI), Lachnospiraceae (cluster XIVa), Ruminococcaceae (cluster IV), Clostridiaceae (cluster I), Erysipelotrichaceae (clusters XVII and XVIII), Peptostreptococciaceae, Unspecified Family XI, Erysipelotrichidae and Bacillarioides, Clostridiaceae, Negatiwickites, Clostridiaceae 2, Clostridiaceae 3 / Halanaerobiales, Veillonellaceae, Acidaminococcaceae, Erysipelotrichidae cluster XVII, Erysipelotrichidae cluster XVIII,Erysipelothrixaceae Cluster XVI, Tenericutes, Erysipelothrixaceae, Fusobacteraceae, Leptotrichaceae, Rhodobacteraceae, Brucellaceae, Rhizobiumaceae, Phyllobacteraceae, Aurantimonadaceae, Brasirhizobiumaceae, Methylobacteraceae, Xanthobacteraceae, Caulobacteraceae, Hyphomicrobiumaceae, Sphingomonasaceae, Acetobacteriaceae, Rhodospirillaceae, Firmicutes, Comamonadaceae, Saterellaceae, Alcaligenesaceae, Burkholderiaceae 1, Oxalobacteraceae, Burkholderiaceae 2, Rhodocyclaceae, Neisseriaceae, Enterobacteriaceae, Pasteurellaceae, Vibriaceae, Aeromonadaceae, Succinivibrionaceae, Moraxellaceae, The phylum is selected from the group consisting of the family Campylobacteria, Pseudomonadaceae, Xanthomonadaceae, Sinobacteria, Epsilon Campylobacteria, Epsilon Helicobacteria, Epsilon Campylobacteria, δ-Desulfovibrioaceae, Vermicobium, Lentispaella, Planctomycetes, Spirochaetes, Deinococcus-Thermus, Synergistes, Archaea / Euryarchaeota, Archaea / Crenarchaeota, Fungi-Ascomycota, Fungi-Basidiomycota, Stramenopiles, Apicomplexa, Amoeba, Ciliata, Metamonada, Microsporidia, Fornicata, Akkermansia, Christenseneraceae, Odoribacteria, Egasellaceae, and combinations thereof.

[0156] In certain embodiments, during step (a), the anaerobic microorganism is selected from the group consisting of Akkermansiaceae, Bacillaceae, Bacteroidetaceae, Bifidobacteriaceae, Christensellaceae, Clostridiaceae, Coriobacteraceae, Egasellaceae, Enterobacteriaceae, Eubacteriaceae, Lachnospiraceae, Lactobacillaceae, Leuconostocaceae, Odoribacteraceae, Oxalobacteraceae, Porphyromonasaceae, Prevotellaceae, Rikenellaceae, Ruminococcaceae, Streptococciaceae, and combinations thereof.

[0157] In certain embodiments, during step (a), the microorganism is in the form of a cell paste. In certain embodiments, the cell paste comprises the microorganism in a cell culture medium. The cell paste can be prepared by cultivating the microorganism and then harvesting the microorganism and, for example, removing the moisture content to form the cell paste.

[0158] In certain embodiments, during step (a), the cell paste is not washed to remove cell culture medium, e.g., with water, a buffer, or a salt solution, prior to combining with the excipient blend.

[0159] In certain embodiments, during step (a), the cryopreservation mixture, when solvated in an aqueous solvent, is from about 30% (w / w) to about 50% (w / w), from about 32% (w / w) to about 50% (w / w), from about 34% (w / w) to about 50% (w / w), from about 36% (w / w) to about 50% (w / w), from about 38% (w / w) to about 50% (w / w), from about 40% (w / w) to about 50% (w / w), from about 42% (w / w) to about 50% (w / w), from about 44% (w / w) to about 50% (w / w), from about 46% (w / w) to about 50% (w / w), from about 48% (w / w) to about 50% (w / w). / w), about 30% (w / w) to about 48% (w / w), about 30% (w / w) to about 46% (w / w), about 30% (w / w) to about 44% (w / w), about 30% (w / w) to about 42% (w / w), about 30% (w / w) to about 40% (w / w), about 30% (w / w) to about 38% (w / w), About 30% (w / w) to about 36% (w / w), about 30% (w / w) to about 34% (w / w), about 30% (w / w) to about 32% (w / w), about 32% (w / w) to about 48% (w / w), about 32% (w / w) to about 46% (w / w), about 32% (w / w) to about 44% (w / w), about 32% (w / w) to about 42% (w / w), about 32% (w / w) to about 40% (w / w), about 32% (w / w) to about 38% (w / w), about 32% (w / w) to about 36% (w / w), about 32% (w / w) to about 34% (w / w), about 34% (w / w) to about 48% (w / w), about 34% (w / w) ) to about 46% (w / w), about 34% (w / w) to about 44% (w / w), about 34% (w / w) to about 42% (w / w), about 34% (w / w) to about 40% (w / w), about 34% (w / w) to about 38% (w / w), about 34% (w / w) to about 36% (w / w), about 36% (w / w) to about 4 8% (w / w), about 36% (w / w) to about 46% (w / w), about 36% (w / w) to about 44% (w / w), about 36% (w / w) to about 42% (w / w), about 36% (w / w) to about 40% (w / w), about 36% (w / w) to about 38% (w / w), about 38% (w / w) to about 48% (w / w), about 38% (w / w) to about 46% (w / w), about 38% (w / w) to about 44% (w / w), about 38% (w / w) to about 42% (w / w), about 38% (w / w) to about 40% (w / w), about 40% (w / w) to about 48% (w / w), about 40% (w / w) to about 46% (w / w),The cell paste or cell slurry may comprise about 40% (w / w) to about 44% (w / w), about 40% (w / w) to about 42% (w / w), about 42% (w / w) to about 48% (w / w), about 42% (w / w) to about 46% (w / w), about 42% (w / w) to about 44% (w / w), about 44% (w / w) to about 48% (w / w), about 44% (w / w) to about 46% (w / w), or about 46% (w / w) to about 48% (w / w). In certain embodiments, during step (a), the cryopreservation mixture comprises about 30% (w / w) to about 50% (w / w) of the cell paste or cell slurry when solvated in an aqueous solvent.

[0160] In certain embodiments, during step (a), the cryopreservation mixture, when solvated in an aqueous solvent, such as water (e.g., ionized or deionized water), is about 30% (w / w), about 31% (w / w), about 32% (w / w), about 33% (w / w), about 34% (w / w), about 35% (w / w), about 36% (w / w), about 37% (w / w), ), about 38% (w / w), about 39% (w / w), about 40% (w / w), about 41% (w / w), about 42% (w / w), about 43% (w / w), about 44% (w / w), about 45% (w / w), about 46% (w / w), about 47% (w / w), about 48% (w / w), about 49% (w / w), or about 50% (w / w) of the cell paste or cell slurry.

[0161] In certain embodiments, during step (a), the aqueous mixture comprises deionized water. In certain embodiments, during step (a), the cryopreservation mixture comprises about 30% (w / w) to about 60% (w / w), about 35% (w / w) to about 60% (w / w), about 40% (w / w) to about 60% (w / w), about 45% (w / w) to about 60% (w / w), about 50% (w / w) to about 60% (w / w), about 55% (w / w) to about 60% (w / w), about 30% (w / w) to about 55% (w / w), about 30% (w / w) to about 50% (w / w), about 30% (w / w) to about 45% (w / w), about 30% (w / w) to about 40% (w / w).

[0043] In one embodiment, the composition comprises about 30% (w / w) to about 35% (w / w), about 35% (w / w) to about 55% (w / w), about 35% (w / w) to about 50% (w / w), about 35% (w / w) to about 45% (w / w), about 35% (w / w) to about 40% (w / w), about 40% (w / w) to about 55% (w / w), about 40% (w / w) to about 50% (w / w), about 40% (w / w) to about 45% (w / w), about 45% (w / w) to about 55% (w / w), about 45% (w / w) to about 50% (w / w), or about 50% (w / w) to about 55% (w / w) of water.

[0162] In certain embodiments, during step (a), the cryopreservation mixture is about 30% (w / w), about 31% (w / w), about 32% (w / w), about 33% (w / w), about 34% (w / w), about 35% (w / w), about 36% (w / w), about 37% (w / w), about 38% (w / w), about 39% (w / w), about 40% (w / w), about 41% (w / w), about 42% (w / w), about 43% (w / w), about 44% (w / w), about 45% (w / w), about 46% (w / w), about 47% (w / w), about 48% (w / w), about 49% (w / w), about 50% (w / w), about 51% (w / w), about 52% (w / w), about 53% (w / w), about 54% (w / w), about 55% (w / w), about 56% (w / w), about 57% (w / w), about 58% (w / w), about 59% (w / w), or about 60% (w / w) water.

[0163] In certain embodiments, during step (a), the cryopreservation mixture is from about 0.01% (w / w) to about 0.4% (w / w), from about 0.05% (w / w) to about 0.4% (w / w), from about 0.1% (w / w) to about 0.4% (w / w), from about 0.15% (w / w) to about 0.4% (w / w), from about 0.2% (w / w) to about 0.4% (w / w), from about 0.25% (w / w) to about 0.4% (w / w), from about 0.3% (w / w) to about 0.4% (w / w), from about 0.35% (w / w) to about 0.4% (w / w), from about 0. 0.01% (w / w) to about 0.35% (w / w), about 0.01% (w / w) to about 0.3% (w / w), about 0.01% (w / w) to about 0.25% (w / w), about 0.01% (w / w) to about 0.2% (w / w), about 0.01% (w / w) to about 0.15% (w / w), about 0.01% (w / w) to about 0.1% (w / w), about 0.01% (w / w) to about 0.05% (w / w), about 0.05% (w / w) to about 0.35% (w / w), about 0.05% (w / w) to about 0.3% (w / w), about 0.05% (w / w) ~ about 0.25% (w / w), about 0.05% (w / w) to about 0.2% (w / w), about 0.05% (w / w) to about 0.15% (w / w), about 0.05% (w / w) to about 0.1% (w / w), about 0.1% (w / w) to about 0.35% (w / w), about 0.1% (w / w) to about 0.3% (w / w), about 0.1% (w / w) to about 0.25% (w / w), about 0.1% (w / w) to about 0.2% (w / w), about 0.1% (w / w) to about 0.15% (w / w), about 0.15% (w / w) to about 0.35% (w / w), The viscosity modifier may be about 0.15% (w / w) to about 0.3% (w / w), about 0.15% (w / w) to about 0.25% (w / w), about 0.15% (w / w) to about 0.2% (w / w), about 0.2% (w / w) to about 0.35% (w / w), about 0.2% (w / w) to about 0.3% (w / w), about 0.2% (w / w) to about 0.25% (w / w), about 0.25% (w / w) to about 0.35% (w / w), about 0.25% (w / w) to about 0.35% (w / w), or about 0.3% (w / w) to about 0.35% (w / w). In certain embodiments, during step (a), the cryopreservation mixture comprises from about 0.01% (w / w) to about 0.4% (w / w) of a viscosity modifier.

[0164] In certain embodiments, during step (a), the cryopreservation mixture is about 0.01% (w / w), about 0.02% (w / w), about 0.03% (w / w), about 0.04% (w / w), about 0.05% (w / w), about 0.06% (w / w), about 0.07% (w / w), about 0.08% (w / w), about 0.09% (w / w), about 0.1% (w / w), about 0.11% (w / w), about 0.12% (w / w), about 0.13% (w / w), about 0.14% (w / w), about 0.15% (w / w), about 0.16% (w / w), about 0.17% (w / w), about 0.18% (w / w), about 0.19% (w / w), , about 0.2% (w / w), about 0.21% (w / w), about 0.22% (w / w), about 0.23% (w / w), about 0.24% (w / w), about 0.25% (w / w), about 0.26% (w / w), about 0.27% (w / w), about 0.28% (w / w), about 0.29% (w / w), about 0.3% (w / w), About 0.31% (w / w), about 0.32% (w / w), about 0.33% (w / w), about 0.34% (w / w), about 0.35% (w / w), about 0.36% (w / w), about 0.37% (w / w), about 0.38% (w / w), about 0.39% (w / w), or about 0.4% (w / w) of a viscosity modifier.

[0165] In certain embodiments, during step (a), the viscosity modifier is a polymer composition, hi certain embodiments, the polymer composition comprises alginate, acacia, alginic acid, agar, aramic acid, bentonite, carbomer, carrageenan, cellulose, dextrin, gelatin, gellan gum, guar gum, carboxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, methylcellulose, mucin, pectin, starch, xanthan gum, or a combination thereof.

[0166] In certain embodiments, the polymer composition comprises an alginate, such as sodium alginate. The is approximately 250cP ~ approximately 14,000cP, approximately 500cP ~ approximately 14,000cP, approximately 750cP ~ approximately 14,000cP, approximately 1,000cP ~ approximately 14,000cP, approximately 2,000cP ~ approximately 14,000cP, approximately 3,000cP ~ approximately 14,000cP, approximately 4,00 0cP ~ approx. 14,000cP, approx. 5,000cP ~ approx. 14,000cP, approx. 6,000cP ~ approx. 14,000cP, approx. 7,000c P ~ approx. 14,000cP, approx. 8,000cP ~ approx. 14,000cP, approx. 9,000cP ~ approx. 14,000cP, approx. 10,000cP ~14,000cP, 11,000cP~14,000cP, 12,000cP~14,000cP, 13,000c P ~ approx. 14,000cP, approx. 250cP ~ approx. 13,000cP, approx. 250cP ~ approx. 12,000cP, approx. 250cP ~ approx. 11,00 0cP, about 250cP to about 10,000cP, about 250cP to about 9,000cP, about 250cP to about 8,000cP, about 250cP ~7,000cP, approximately 250cP~6,000cP, approximately 250cP~5,000cP, approximately 250cP~4,000cP, approximately 2 50cP ~ approx. 3,000cP, approx. 250cP ~ approx. 2,000cP, approx. 250cP ~ approx. 1,000cP, approx. 250cP ~ approx. 750cP , about 250cP to about 500cP, about 500cP to about 13,000cP, about 500cP to about 12,000cP, about 500cP to about 11 ,000cP, about 500cP to about 10,000cP, about 500cP to about 9,000cP, about 500cP to about 8,000cP, about 50 0cP ~ approx. 7,000cP, approx. 500cP ~ approx. 6,000cP, approx. 500cP ~ approx. 5,000cP, approx. 500cP ~ approx. 4,000cP , about 500cP to about 3,000cP, about 500cP to about 2,000cP, about 500cP to about 1,000cP, about 500cP to about 75 0cP, about 750cP to about 13,000cP, about 750cP to about 12,000cP, about 750cP to about 11,000cP, about 750 cP ~ approx. 10,000cP, approx. 750cP ~ approx. 9,000cP, approx. 750cP ~ approx. 8,000cP, approx. 750cP ~ approx. 7,000c P, about 750cP to about 6,000cP, about 750cP to about 5,000cP, about 750cP to about 4,000cP, about 750cP to about 3,000 cP, about 750 cP to about 2,000 cP, about 750 cP to about 1,000 cP, about 1,000 cP to about 13,000 cP, about 1,000 cP to about 12,000 cP, about 1,000 cP to about 11,000 cP, about 1,000 cP to about 10,000 cP, about 1,000 cP to about 9,000 cP, about 1,000 cP to about 8,000 cP, about 1,000 cP to about 7,000 cP, about 1,000 cP to about 6,000 cP, about 1,000 cP to about 5,000 cP, about 1,000 cP to about 4,000 cP, about 1,000 cP to about 3,000 cP, about 1,000 cP to about 2,000 cP, about 2,000 cP to about 13,000 cP, about 2,000 cP to about 12,000 cP, about 2,000 cP to about 11,000 cP, about 2,000 cP to about 10,000 cP, about 2,000 cP to about 9,000 cP, about 2,000 cP to about 8,000 cP, about 2,000 cP to about 7,000 cP, about 2,000 cP to about 6,000 cP, about 2,000 cP to about 5,000 cP, about 2,000 cP to about 4,000 cP, about 2,000 cP to about 3,000 cP, about 3,000 cP to about 13,000 cP, about 3,000 cP to about 12,000 cP, about 3,000 cP to about 11,000 cP, about 3,000 cP to about 10,000 cP, about 3,000 cP to about 9,000 cP, about 3,000 cP to about 8,000 cP, about 3,000 cP to about 7,000 cP, about 3,000 cP to about 6,000 cP, about 3,000 cP to about 5,000 cP, about 3,000 cP to about 4,000 cP, about 4,000 cP to about 13,000 cP, about 4,000 cP to about 12,000 cP, about 4,000 cP to about 11,000 cP, about 4,000 cP to about 10,000 cP, about 4,000 cP to about 9,000 cP, about 4,000 cP to about 8,000 cP, about 4,000 cP to about 7,000 cP, about 4,000 cP to about 6,000 cP, about 4,000 cP to about 5,000 cP, about 5,000 cP to about 13,000 cP, about 5,000 cP to about 12,000 cP, about 5,000 cP to about 11,000 cP, about 5,000 cP to about 10,000 cP, about 5,000 cP to about 9,000 cP, about 5,000 cP to about 8,000 cP, about 5,000 cP to about 7,000 cP, about 5,000 cP to about 6,000 cP, about 6,000 cP to about 13,000 cP, about 6,000 cP to about 12,000 cP, about 6,000cP ~ approx. 11,000cP, approx. 6,000cP ~ approx. 10,000cP, approx. 6,000cP ~ approx. 9,000cP, approx. 6,000cP ~ approx. 8,000cP, approximately 6,000cP to approximately 7,000cP, approximately 7,000cP to approximately 13,000cP, approximately 7,000cP to approximately 12,000cP , about 7,000cP to about 11,000cP, about 7,000cP to about 10,000cP, about 7,000cP to about 9,000cP, about 7,000 cP ~ approx. 8,000cP, approx. 8,000cP ~ approx. 13,000cP, approx. 8,000cP ~ approx. 12,000cP, approx. 8,000cP ~ approx. 11, 000 cP, about 8,000 cP to about 10,000 cP, about 8,000 cP to about 9,000 cP, about 9,000 cP to about 13,000 cP, about 9,000 cP to about 12,000 cP, about 9,000 cP to about 11,000 cP, about 9,000 cP to about 10,000 cP, about 10,000 cP to about 13,000 cP, about 10,000 cP to about 12,000 cP, about 10,000 cP to about 11,000 cP, about 11,000 cP to about 13,000 cP, about 11,000 cP to about 12,000 cP, or about 12,000 cP to about 13,000 cP. In certain embodiments, sodium alginate has a viscosity range of about 250 cP to about 14,000 cP.

[0167] In certain embodiments, during step (a), the cryopreservation mixture is from about 3% (w / w) to about 15% (w / w), from about 5% (w / w) to about 15% (w / w), from about 7% (w / w) to about 15% (w / w), from about 9% (w / w) to about 15% (w / w), from about 11% (w / w) to about 15% (w / w), from about 13% (w / w) to about 15% (w / w), from about 3% (w / w) to about 13% (w / w), from about 3% (w / w) to about 11% (w / w), from about 3% (w / w) to about 9% (w / w), from about 3% (w / w) to about 7% (w / w),

[0043] The energy source may comprise about 3% (w / w) to about 5% (w / w), about 5% (w / w) to about 13% (w / w), about 5% (w / w) to about 11% (w / w), about 5% (w / w) to about 9% (w / w), about 5% (w / w) to about 7% (w / w), about 7% (w / w) to about 13% (w / w), about 7% (w / w) to about 11% (w / w), about 7% (w / w) to about 9% (w / w), about 9% (w / w) to about 13% (w / w), about 9% (w / w) to about 11% (w / w), or about 11% (w / w) to about 13% (w / w). In certain embodiments, during step (a), the cryopreservation mixture comprises from about 3% (w / w) to about 15% (w / w) of the energy source.

[0168] In certain embodiments, during step (a), the cryopreservation mixture comprises about 3% (w / w), about 4% (w / w), about 5% (w / w), about 6% (w / w), about 7% (w / w), about 8% (w / w), about 9% (w / w), about 10% (w / w), about 11% (w / w), about 12% (w / w), about 13% (w / w), about 14% (w / w), or about 15% (w / w) of the energy source.

[0169] In certain embodiments, during step (a), the energy source is a monosaccharide (e.g., glucose, fructose, xylose, and combinations thereof), a disaccharide (e.g., trehalose, sucrose, or combinations thereof), an oligosaccharide (e.g., maltodextrin or an inulin derivative (e.g., fructooligosaccharide such as raftilose), or a combination thereof. In certain embodiments, during step (a), the energy source comprises raftilose and maltodextrin.

[0170] In certain embodiments, during step (a), the cryopreservation mixture is from about 0.5% (w / w) to about 5% (w / w), from about 1% (w / w) to about 5% (w / w), from about 1.5% (w / w) to about 5% (w / w), from about 2% (w / w) to about 5% (w / w), from about 2.5% (w / w) to about 5% (w / w), from about 3% (w / w) to about 5% (w / w), from about 3.5% (w / w) to about 5% (w / w), from about 4% (w / w) to about 5% (w / w), from about 4.5% (w / w) to about 5% (w / w), from about 0.5% (w / w) to about 4.5% (w / w), or from about 0.5% (w / w). (w / w) to about 4% (w / w), about 0.5% (w / w) to about 3.5% (w / w), about 0.5% (w / w) to about 3% (w / w), about 0.5% (w / w) to about 2.5% (w / w), about 0.5% (w / w) to about 2% (w / w), about 0.5% (w / w) to about 1.5% (w / w), about 0.5% (w / w) to about 1% (w / w), about 1% (w / w) to about 4.5% (w / w), about 1% (w / w) to about 4% (w / w), about 1% (w / w) to about 3.5% (w / w), about 1% (w / w) to about 3% (w / w), about 1% (w / w) to about 2.5% (w / w), about 1% (w / w) to about 2% (w / w), about 1% (w / w) to about 1.5% (w / w), about 1.5% (w / w) to about 4.5% (w / w), about 1.5% (w / w) to about 4% (w / w), about 1.5% (w / w) to about 3.5% (w / w), about 1.5% (w / w) to about 3% (w / w), about 1.5% (w / w) to about 2.5% (w / w), about 1.5% (w / w) to about 2% (w / w), about 2% (w / w) to about 4.5% (w / w), about 2% (w / w) to about 4% (w / w), about 2% (w / w) to about 3.5% (w / w), about 2% (w / w) to about 3% (w / w), The composition contains about 2% (w / w) to about 2.5% (w / w), about 2.5% (w / w) to about 4.5% (w / w), about 2.5% (w / w) to about 4% (w / w), about 2.5% (w / w) to about 3.5% (w / w), about 2.5% (w / w) to about 3% (w / w), about 3% (w / w) to about 4.5% (w / w), about 3% (w / w) to about 4% (w / w), about 3% (w / w) to about 3.5% (w / w), about 3.5% (w / w) to about 4.5% (w / w), about 3.5% (w / w) to about 4% (w / w), or about 4% (w / w) to about 4.5% (w / w).In certain embodiments, during step (a), the cryopreservation mixture comprises about 0.5% (w / w) to about 5% (w / w) of raftilose.

[0171] In certain embodiments, during step (a), the cryopreservation composition comprises about 0.5% (w / w), about 1% (w / w), about 1.5% (w / w), about 2% (w / w), about 3% (w / w), about 3.5% (w / w), about 4% (w / w), about 4.5% (w / w), or about 5% (w / w) of raftilose.

[0172] In certain embodiments, during step (a), the cryopreservation mixture is from about 3% (w / w) to about 10% (w / w), from about 4% (w / w) to about 10% (w / w), from about 5% (w / w) to about 10% (w / w), from about 6% (w / w) to about 10% (w / w), from about 7% (w / w) to about 10% (w / w), from about 8% (w / w) to about 10% (w / w), about 9% (w / w) to about 10% (w / w), about 3% (w / w) to about 9% (w / w), about 3% (w / w) to about 8% (w / w), about 3% (w / w) to about 7% (w / w), about 3% (w / w) to about 6% (w / w), about 3% (w / w) to about 5% (w / w), about 3% (w / w) to about 4% (w / w), about 4% (w / w) to about 9% (w / w), about 4% (w / w) to about 8% (w / w), about 4% (w / w) to about 7% (w / w), about 4% (w / w) to about 6% (w / w), about 4% (w / w) to about 5% (w / w), about 5% (w / w) to about 9% (w / w), about 5% (w / w) to about 8% (w / w), about 5% (w / w) to about 7% (w / w), about The mixture may comprise 5% (w / w) to about 6% (w / w), about 6% (w / w) to about 9% (w / w), about 6% (w / w) to about 8% (w / w), about 6% (w / w) to about 7% (w / w), about 7% (w / w) to about 9% (w / w), about 7% (w / w) to about 8% (w / w), or about 8% (w / w) to about 9% (w / w) of maltodextrin. In certain embodiments, during step (a), the cryopreservation mixture comprises about 3% (w / w) to about 10% (w / w) of maltodextrin.

[0173] In certain embodiments, during step (a), the cryopreservation mixture contains about 3% (w / w), about 3.5% (w / w), about 4% (w / w), about 4.5% (w / w), about 5% (w / w), about 5.5% (w / w), about 6% (w / w), about 6.5% (w / w), about 7% (w / w), about 7.5% (w / w), about 8% (w / w), about 8.5% (w / w), about 9% (w / w), about 9.5% (w / w), or about 10% (w / w) maltodextrin.

[0174] In certain embodiments, during step (a), the aqueous mixture further comprises a protein stabilizing agent. In certain embodiments, during step (a), the cryopreservation mixture is about 5% (w / w) to about 10% (w / w), about 6% (w / w) to about 10% (w / w), about 7% (w / w) to about 10% (w / w), about 8% (w / w) to about 10% (w / w), about 9% (w / w) to about 10% (w / w), about 5% (w / w) to about 9% (w / w), about 5% (w / w) to about 8% (w / w), about 10 ...

[0043] In some embodiments, the composition comprises about 5% (w / w) to about 7% (w / w), about 5% (w / w) to about 6% (w / w), about 6% (w / w) to about 9% (w / w), about 6% (w / w) to about 8% (w / w), about 6% (w / w) to about 7% (w / w), about 7% (w / w) to about 9% (w / w), about 7% (w / w) to about 8% (w / w), or about 8% (w / w) to about 9% (w / w) of the protein stabilizer.

[0175] In certain embodiments, during step (a), the cryopreservation mixture comprises about 5% (w / w), about 5.5% (w / w), about 6% (w / w), about 6.5% (w / w), about 7% (w / w), about 7.5% (w / w), about 8% (w / w), about 8.5% (w / w), about 9% (w / w), about 9.5% (w / w), or about 10% (w / w) of a protein stabilizing agent.

[0176] In certain embodiments, the protein stabilizing agent can be selected from the group consisting of amino acids, antioxidants, disaccharides, reducing agents, polyethylene glycol, and combinations thereof. The disaccharides can be selected from the group consisting of trehalose, sucrose, isomaltose, lactose, maltose, lactulose, cellobiose, and combinations thereof. In certain embodiments, the protein stabilizing agent comprises trehalose. In certain embodiments, the protein stabilizing agent comprises sucrose and trehalose.

[0177] In certain embodiments, during step (a), the cryopreservation mixture is from about 5% (w / w) to about 10% (w / w), from about 6% (w / w) to about 10% (w / w), from about 7% (w / w) to about 10% (w / w), from about 8% (w / w) to about 10% (w / w), from about 9% (w / w) to about 10% (w / w), from about 5% (w / w) to about 9% (w / w), from about 5% (w / w) to about 8% (w / w), In certain embodiments, during step (a), the cryopreservation mixture comprises from about 5% (w / w) to about 7% (w / w), from about 5% (w / w) to about 6% (w / w), from about 6% (w / w) to about 9% (w / w), from about 6% (w / w) to about 8% (w / w), from about 6% (w / w) to about 7% (w / w), from about 7% (w / w) to about 9% (w / w), from about 7% (w / w) to about 8% (w / w), or from about 8% (w / w) to about 9% (w / w). In certain embodiments, during step (a), the cryopreservation mixture comprises from about 5% (w / w) to about 10% (w / w) trehalose.

[0178] In certain embodiments, during step (a), the cryopreservation mixture comprises about 5% (w / w), about 5.5% (w / w), about 6% (w / w), about 6.5% (w / w), about 7% (w / w), about 7.5% (w / w), about 8% (w / w), about 8.5% (w / w), about 9% (w / w), about 9.5% (w / w), or about 10% (w / w) trehalose.

[0179] In certain embodiments, during step (a), the cryopreservation mixture is from 0% (w / w) to about 10% (w / w), from about 1% (w / w) to about 10% (w / w), from about 2% (w / w) to about 10% (w / w), from about 3% (w / w) to about 10% (w / w), from about 4% (w / w) to about 10% (w / w), from about 5% (w / w) to about 10% (w / w), from about 6% (w / w) to about 10% (w / w), from about 7% (w / w) to about 10% (w / w), from about 8% (w / w) to about 10% (w / w), from about 9% (w / w) to about 10% (w / w), 0% (w / w) to about 9% (w / w), 0% (w / w) ) ~ approx. 8% (w / w), 0% (w / w) ~ approx. 7% (w / w), 0% (w / w) ~ approx. 6% (w / w), 0% (w / w) ~ approx. 5% (w / w) , 0% (w / w) ~ approx. 4% (w / w), 0% (w / w) ~ approx. 3% (w / w), 0% (w / w) ~ approx. 2% (w / w), 0% (w / w) ~ approx. 1 %(w / w), about 1%(w / w) to about 9%(w / w), about 1%(w / w) to about 8%(w / w), about 1%(w / w) to about 7%(w / w), about 1%(w / w) to about 6%(w / w), about 1%(w / w) to about 5%(w / w), about 1%(w / w) to about 4%(w / w), about 1%(w / w) to About 3% (w / w), about 1% (w / w) to about 2% (w / w), about 2% (w / w) to about 9% (w / w), about 2% (w / w) to about 8% (w / w), about 2% (w / w) to about 7% (w / w), about 2% (w / w) to about 6% (w / w), about 2% (w / w) to about 5% (w / w), about 2% (w / 3%(w / w) to about 4%(w / w), about 2%(w / w) to about 3%(w / w), about 3%(w / w) to about 9%(w / w), about 3%(w / w) to about 8%(w / w), about 3%(w / w) to about 7%(w / w), about 3%(w / w) to about 6%(w / w), about 3%(w / w) to about 5%(w / w), about 3%( (w / w) to about 4% (w / w), about 4% (w / w) to about 9% (w / w), about 4% (w / w) to about 8% (w / w), about 4% (w / w) to about 7% (w / w), about 4% (w / w) to about 6% (w / w), about 4% (w / w) to about 5% (w / w), about 5% (w / w) to about 9% (w / w), about 5 %(w / w) to about 8%(w / w), about 5%(w / w) to about 7%(w / w), about 5%(w / w) to about 6%(w / w), about 6%(w / w) to about 9%(w / w), about 6%(w / w) to about 8%(w / w), about 6%(w / w) to about 7%(w / w), about 7%(w / w) to about 9%(w / w),About 7% (w / w) to about 8% (w / w), or about 8% (w / w) to about 9% (w / w) sucrose. In certain embodiments, during step (a), the cryopreservation mixture, when solvated in an aqueous solvent, comprises about 5% (w / w) to about 10% (w / w) sucrose. In certain embodiments, during step (a), the cryopreservation mixture comprises 0% (w / w) to about 5% (w / w) sucrose.

[0180] In certain embodiments, during step (a), the cryopreservation mixture comprises about 0.5% (w / w), about 1% (w / w), about 1.5% (w / w), about 2% (w / w), about 2.5% (w / w), about 3% (w / w), about 3.5% (w / w), about 4% (w / w), about 4.5% (w / w), about 5% (w / w), about 5.5% (w / w), about 6% (w / w), about 6.5% (w / w), about 7% (w / w), about 7.5% (w / w), about 8% (w / w), about 8.5% (w / w), about 9% (w / w), about 9.5% (w / w), or about 10% (w / w) sucrose.

[0181] In certain embodiments, during step (a), the aqueous mixture further comprises a buffering agent, such as a sodium phosphate buffer system or a potassium phosphate buffer system.

[0182] In certain embodiments, during step (a), the cryopreservation mixture has a concentration of from about 5 cP to about 6,000 cP, from about 50 cP to about 6,000 cP, from about 100 cP to about 6,000 cP, from about 200 cP to about 6,000 cP, from about 300 cP to about 6,000 cP, from about 400 cP to about 6,000 cP, from about 500 cP to about 6,000 cP, from about 1,000 cP to about 6,000 cP, from about 1,500 cP to about 6,000 cP, from about 2,000 cP to about 6,000 cP, from about 2,500 cP to about 6,000 cP, from about 3,000 cP to about 6,000 cP, from about 3,500 cP to about 6,0 00cP, approx. 4,000cP ~ approx. 6,000cP, approx. 4,500cP ~ approx. 6,000cP, approx. 5,000cP ~ approx. 6,000cP, approx. 5, 500cP~Approx. 6,000cP, Approx. 5cP~Approx. 5,500cP, Approx. 5cP~Approx. 5,000cP, Approx. 5cP~Approx. 5,000cP, Approx. 5cP~ Approx. 4,500cP, Approx. 5cP ~ Approx. 4,000cP, Approx. 5cP ~ Approx. 3,500cP, Approx. 5cP ~ Approx. 3,000cP, Approx. 5cP ~ Approx. 2,500 cP, about 5 cP to about 2,000 cP, about 5 cP to about 1,500 cP, about 5 cP to about 1,000 cP, about 5 cP to about 500 cP, about 5 cP to Approximately 400cP, approximately 5cP to approximately 300cP, approximately 5cP to approximately 200cP, approximately 5cP to approximately 100cP, approximately 5cP to approximately 50cP, approximately 50cP to approximately 5,500cP, about 50cP to about 5,000cP, about 50cP to about 4,500cP, about 50cP to about 4,000cP, about 50cP to about 3, 500cP, about 50cP to about 3,000cP, about 50cP to about 2,500cP, about 50cP to about 2,000cP, about 50cP to about 1,50 0cP, about 50cP to about 1,000cP, about 50cP to about 500cP, about 50cP to about 400cP, about 50cP to about 300cP, about 50c P ~ about 200cP, about 50cP - about 100cP, about 100cP - about 5,500cP, about 100cP - about 5,000cP, about 100cP - about 4,500cP, about 100cP to about 4,000cP, about 100cP to about 3,500cP, about 100cP to about 3,000cP, about 100cP ~2,500cP, 100cP~2,000cP, 100cP~1,500cP, 100cP~1,000cP, 100 cP ~ approx. 500cP, approx. 100cP ~ approx. 400cP, approx. 100cP ~ approx. 300cP, approx. 100cP ~ approx. 200cP, approx. 200cP ~ approx. 5,500 cP, from about 200 cP to about 5,000 cP, from about 200 cP to about 4,500 cP, from about 200 cP to about 4,000 cP, from about 200 cP to about 3,500 cP, from about 200 cP to about 3,000 cP, from about 200 cP to about 2,500 cP, from about 200 cP to about 2,000 cP, from about 200 cP to about 1,500 cP, from about 200 cP to about 1,000 cP, from about 200 cP to about 500 cP, from about 200 cP to about 400 cP, from about 200 cP to about 300 cP, from about 300 cP to about 5,500 cP, from about 300 cP to about 5,000 cP, from about 300 cP to about 4,500 cP, from about 300 cP to about 4,000 cP, from about 300 cP to about 3,500 cP, from about 300 cP to about 3,000 cP, from about 300 cP to about 2,500 cP, from about 300 cP to about 2,000 cP, from about 300 cP to about 1,500 cP, from about 300 cP to about 1,000 cP, from about 300 cP to about 500 cP, from about 300 cP to about 400 cP, from about 400 cP to about 5,500 cP, from about 400 cP to about 5,000 cP, from about 400 cP to about 4,500 cP, from about 400 cP to about 4,000 cP, from about 400 cP to about 3,500 cP, from about 400 cP to about 3,000 cP, from about 400 cP to about 2,500 cP, from about 400 cP to about 2,000 cP, from about 400 cP to about 1,500 cP, from about 400 cP to about 1,000 cP, from about 400 cP to about 500 cP, from about 500 cP to about 5,500 cP, from about 500 cP to about 5,000 cP, from about 500 cP to about 4,500 cP, from about 500 cP to about 4,000 cP, from about 500 cP to about 3,500 cP, from about 500 cP to about 3,000 cP, from about 500 cP to about 2,500 cP, from about 500 cP to about 2,000 cP, from about 500 cP to about 1,500 cP, from about 500 cP to about 1,000 cP, from about 1,000 cP to about 5,500 cP, from about 1,000 cP to about 5,000 cP, from about 1,000 cP to about 4,500 cP, from about 1,000 cP to about 4,000 cP, from about 1,000 cP to about 3,500 cP, from about 1,000 cP to about 3,000 cP, from about 1,000 cP to about 2,500 cP, from about 1,000 cP to about 2,000 cP, from about 1,000 cP to about 1,500 cP, from about 1,500 cP to about 5,500 cP, from about 1,500 cP to about 5,000 cP, from about 1,500 cP to about 4,500 cP, from about 1,500 cP to about 4,000 cP, from about 1,500 cP to about 3,500 cP, from about 1,500 cP to about 3,000 cP, from about 1,500 cP to about 2,500cP, approximately 1,500cP to approximately 2,000cP, approximately 2,000cP to approximately 5,500cP, approximately 2,000cP to approximately 5,000cP, approximately 2,000c P ~ about 4,500cP, about 2,000cP - about 4,000cP, about 2,000cP - about 3,500cP, about 2,000cP - about 3,000cP, about 2 ,000cP ~ approx. 2,500cP, approx. 2,500cP ~ approx. 5,500cP, approx. 2,500cP ~ approx. 5,000cP, approx. 2,500cP ~ approx. 4,500 cP, approximately 2,500cP to approximately 4,000cP, approximately 2,500cP to approximately 3,500cP, approximately 2,500cP to approximately 3,000cP, approximately 3,000cP to approximately 5 ,500cP, approx. 3,000cP ~ approx. 5,000cP, approx. 3,000cP ~ approx. 4,500cP, approx. 3,000cP ~ approx. 4,000cP, approx. 3,000 cP ~ approx. 3,500cP, approx. 3,500cP ~ approx. 5,500cP, approx. 3,500cP ~ approx. 5,000cP, approx. 3,500cP ~ approx. 4,500cP, approx. It has a viscosity in the range of 3,500 cP to about 4,000 cP, about 4,000 cP to about 5,500 cP, about 4,000 cP to about 5,000 cP, about 4,000 cP to about 4,500 cP, about 4,500 cP to about 5,500 cP, about 4,500 cP to about 5,000 cP, about 5,000 cP to about 5,500 cP.

[0183] In certain embodiments, during step (a), the cryopreservation mixture has a viscosity in the range of about 50 cP to about 4,000 cP, about 100 cP to about 2,000 cP, or about 100 cP to 1,000 cP. In certain embodiments, during step (a), the cryopreservation mixture has a viscosity in the range of about 100 cP to about 1,000 cP.

[0184] In certain embodiments, during step (a), the cryopreservation mixture has a concentration of about 5 cP, about 10 cP, about 15 cP, about 20 cP, about 25 cP, about 30 cP, about 35 cP, about 40 cP, about 45 cP, about 50 cP, about 55 cP, about 60 cP, about 65 cP, about 70 cP, about 75 cP, about 80 cP, about 85 cP, about 90 cP, about 95 cP, about 100 cP, about 110 cP, about 120 cP, about 130 cP, about 140 cP, about 150 cP, about 160 cP, about 170 cP, about 180 cP, about The viscosity is about 190 cP, about 200 cP, about 225 cP, about 250 cP, about 275 cP, about 300 cP, about 350 cP, about 400 cP, about 450 cP, about 500 cP, about 600 cP, about 700 cP, about 800 cP, about 900 cP, about 1,000 cP, about 1,250 cP, about 1,500 cP, about 2,000 cP, about 2,500 cP, about 3,000 cP, about 3,500 cP, about 4,000 cP, about 4,500 cP, about 5,000 cP, about 5,500 cP, or about 6,000 cP.

[0185] In certain embodiments, during step (a), the cryopreservation mixture is from about 300 Osm / L to about 4,000 Osm / L, from about 400 Osm / L to about 4,000 Osm / L, from about 500 Osm / L to about 4,000 Osm / L, from about 600 Osm / L to about 4,000 Osm / L, from about 700 Osm / L to about 4,000 Osm / L, from about 800 Osm / L to about 4,000 Osm / L, from about 900 Osm / L to about 4,000 Osm / L, from about 1,000 Osm / L to about 4,000 Osm / L, from about 1,500 Osm / L to about 4,000 Osm / L, from about 2,000 Osm / L to about 4,000 Osm / L, / L ~ approx. 4,000Osm / L, approx. 2,500Osm / L ~ approx. 4,000Osm / L, approx. 3,000Osm / L ~ approx. 4,000Osm / L, about 3,500Osm / L~about 4,000Osm / L, about 300Osm / L~about 3,500Osm / L, about 300Osm / L~about 3,0 00Osm / L, about 300Osm / L to about 2,500Osm / L, about 300Osm / L to about 2,000Osm / L, about 300Osm / L ~Approx. 1,500Osm / L, Approx. 300Osm / L ~ Approx. 1,000Osm / L, Approx. 300Osm / L ~ Approx. 900Osm / L, Approx. 300Osm / L ~ Approx. 800Osm / L, Approx. 300Osm / L ~ Approx. 700Osm / L, Approx. 300Osm / L ~ Approx. 600Osm / L, Approx. 300Osm / L ~ approx. 500Osm / L, approx. 300Osm / L ~ approx. 400Osm / L, approx. 400Osm / L ~ approx. 3,500Osm / L, approx. 400Osm / L ~ approx. 3,000Osm / L, approx. 400Osm / L ~ approx. 2,500Osm / L, approx. 400Osm / L ~ approx. 2,500Osm / L, approx. 40 0Osm / L~Approx. 2,000Osm / L, Approx. 400Osm / L~Approx. 2,000Osm / L, Approx. 400Osm / L~Approx. 1,500Osm / L , about 400Osm / L~about 1,000Osm / L, about 400Osm / L~about 900Osm / L, about 400Osm / L~about 800Osm / L, about 400Osm / L to about 700Osm / L, about 400Osm / L to about 600Osm / L, about 400Osm / L to about 500Osm / L, Approx. 500Osm / L ~ Approx. 3,500Osm / L, Approx. 500Osm / L ~ Approx. 3,000Osm / L, Approx. 500Osm / L ~ Approx. 2,500O sm / L, about 500Osm / L to about 2,000Osm / L, about 500Osm / L to about 1,500Osm / L, about 500Osm / L to about 1,000 Osm / L, from about 500 Osm / L to about 900 Osm / L, from about 500 Osm / L to about 800 Osm / L, from about 500 Osm / L to about 700 Osm / L, from about 500 Osm / L to about 600 Osm / L, from about 600 Osm / L to about 3,500 Osm / L, from about 600 Osm / L to about 3,000 Osm / L, from about 600 Osm / L to about 2,500 Osm / L, from about 600 Osm / L to about 2,000 Osm / L, from about 600 Osm / L to about 1,500 Osm / L, from about 600 Osm / L to about 1,000 Osm / L, from about 600 Osm / L to about 900 Osm / L, from about 600 Osm / L to about 800 Osm / L, from about 600 Osm / L to about 700 Osm / L, from about 700 Osm / L to about 3,500 Osm / L, from about 700 Osm / L to about 3,000 Osm / L, from about 700 Osm / L to about 2,500 Osm / L, from about 700 Osm / L to about 2,000 Osm / L, from about 700 Osm / L to about 1,500 Osm / L, from about 700 Osm / L to about 1,000 Osm / L, from about 700 Osm / L to about 900 Osm / L, from about 700 Osm / L to about 800 Osm / L, from about 800 Osm / L to about 3,500 Osm / L, from about 800 Osm / L to about 3,000 Osm / L, from about 800 Osm / L to about 2,500 Osm / L, from about 800 Osm / L to about 2,000 Osm / L, from about 800 Osm / L to about 1,500 Osm / L, from about 800 Osm / L to about 1,000 Osm / L, from about 800 Osm / L to about 900 Osm / L, from about 900 Osm / L to about 3,500 Osm / L, from about 900 Osm / L to about 3,000 Osm / L, from about 900 Osm / L to about 2,500 Osm / L, from about 900 Osm / L to about 2,000 Osm / L, from about 900 Osm / L to about 1,500 Osm / L, from about 900 Osm / L to about 1,000 Osm / L, from about 1,000 Osm / L to about 3,500 Osm / L, from about 1,000 Osm / L to about 3,000 Osm / L, from about 1,000 Osm / L to about 2,500 Osm / L, from about 1,000 Osm / L to about 2,000 Osm / L, from about 1,000 Osm / L to about 1,500 Osm / L, from about 1,500 Osm / L to about 3,500 Osm / L, from about 1,500 Osm / L to about 3,000 Osm / L, from about 1,500 Osm / L to about 2,500 Osm / L, from about 1,500 Osm / L to about 2,000 Osm / L, from about 2,000 Osm / L to about 3,500 Osm / L, from about 2,000 Osm / L to about 3,The osmolality ranges from about 2,000 Osm / L to about 2,500 Osm / L, about 2,500 Osm / L to about 3,500 Osm / L, about 2,500 Osm / L to about 3,000 Osm / L, or about 3,000 Osm / L to about 3,500 Osm / L.

[0186] In certain embodiments, during step (a), the cryopreservation mixture has an osmolality ranging from about 300 Osm / L to about 3,000 Osm / L, from about 400 Osm / L to about 3,000 Osm / L, or from about 500 Osm / L to about 3,000 Osm / L. In certain embodiments, during step (a), the cryopreservation mixture has an osmolality ranging from about 500 Osm / L to about 3,000 Osm / L.

[0187] In certain embodiments, during step (a), the cryopreservation mixture is at about 300 Osm / L, about 350 Osm / L, about 400 Osm / L, about 450 Osm / L, about 500 Osm / L, about 600 Osm / L, about 700 Osm / L, about 800 Osm / L, about 900 Osm / L, about 1,000 Osm / L, having an osmolality of about 1,250 Osm / L, about 1,500 Osm / L, about 1,750 Osm / L, about 2,000 Osm / L, about 2,250 Osm / L, about 2,500 Osm / L, about 2,750 Osm / L, about 3,000 Osm / L, about 3,500 Osm / L, or about 4,000 Osm / L.

[0188] In certain embodiments, during step (a), the cryopreservation mixture is from about 4 to about 8, from about 4.5 to about 8, from about 5 to about 8, from about 5.5 to about 8, from about 6 to about 8, from about 6.5 to about 8, from about 7 to about 8, from about 7.5 to about 8, from about 4 to about 7.5, from about 4 to about 7, from about 4 to about 6.5, from about 4 to about 6, from about 4 to about 5.5, from about 4 to about 5, from about 4 to about 4.5, from about 4.5 to about 7.5, from about 4.5 to about 7, from about 4.5 to about 7.5 The pH range is about 6.5, about 4.5 to about 6, about 4.5 to about 5.5, about 4.5 to about 5, about 5 to about 7.5, about 5 to about 7, about 5 to about 6.5, about 5 to about 6, about 5 to about 5.5, about 5.5 to about 7.5, about 5.5 to about 7, about 5.5 to about 6.5, about 5.5 to about 6, about 6 to about 7.5, about 6 to about 7, about 6 to about 6.5, about 6.5 to about 7.5, about 6.5 to about 7, or about 7 to about 7.5.

[0189] In certain embodiments, during step (a), the cryopreservation mixture has a pH in the range of about 4 to about 8, about 5 to about 8, about 4 to about 7.5, about 5 to about 7.5, or about 5.5 to about 7.5. In certain embodiments, during step (a), the cryopreservation composition has a pH in the range of about 5.5 to about 7.5.

[0190] In certain embodiments, during step (a), the cryopreservation mixture has a pH of about 4, about 4.1, about 4.2, about 4.3, about 4.4, about 4.5, about 4.6, about 4.7, about 4.8, about 4.9, about 5, about 5.1, about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, about 6, about 6.1, about 6.2, about 6.3, about 6.4, about 6.5, about 6.6, about 6.7, about 6.8, about 6.9, about 7, about 7.1, about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7, about 7.8, about 7.9, or about 8.

[0191] In certain embodiments, during step (a), the cryopreservation mixture is at any of the following voltages: -50mV to -500mV, -50mV to -450mV, -50mV to -400mV, -50mV to -350mV, -50mV to -300mV, -50mV to -250mV, -50mV to -200mV, -50mV to -150mV, -50mV to -100mV, -100mV to -500mV, -1 00mV~-450mV, -100mV~-400mV, -100mV~-350mV, -100mV~-300mV, -100mV~-250mV, -100mV~-200mV, -1 00mV~-150mV, -150mV~-500mV, -150mV~-450mV, -150mV~-400mV, -150mV~-350mV, -150mV~-300mV, -1 50mV~-250mV, -150mV~-200mV, -200mV~-500mV, -200mV~-450mV, -200mV~-400mV, -200mV~-350mV, -2 00mV~-300mV, -200mV~-250mV, -250mV~-500mV, -250mV~-450mV, -250mV~-400mV, -250mV~-350mV, -2 The cryopreservation mixture has a redox potential in the range of 50mV to -300mV, -300mV to -500mV, -300mV to -450mV, -300mV to -400mV, -300mV to -350mV, -350mV to -500mV, -350mV to -450mV, -350mV to -400mV, -400mV to -500mV, -400mV to -450mV, or -450mV to -500mV. In certain embodiments, during step (a), the cryopreservation mixture has a redox potential in the range of -50mV to -500mV when solvated in an aqueous solvent.

[0192] In certain embodiments, during step (a), the cryopreservation mixture is at about -50mV, about -60mV, about -70mV, about -80mV, about -90mV, about -100mV, about -110mV, about -120mV, about -130mV, about -140mV, about -150mV, about -160mV, about -170mV, about -180mV, has a redox potential of about -190mV, about -200mV, about -240mV, about -260mV, about -280mV, about -300mV, about -320mV, about -340mV, about -360mV, about -380mV, about -400mV, about -420mV, about -440mV, about -460mV, about -480mV, or about -500mV.

[0193] In certain embodiments, the cryopreservation method further comprises freezing the cryopreservation mixture prior to step (b). It will be appreciated that the freezing process should be tailored to the organism or group of organisms to be cryopreserved. In certain embodiments, the cryopreservation mixture can contain a cryoprotectant and / or a lyoprotectant. Depending on the desired organism, the cell composition can be subjected to various freezing rates. The resulting mixture can then be tested to determine the appropriate freezing rate for the microorganism or group of microorganisms. It will be appreciated that when a group of microorganisms is frozen together, the selected freezing rate can be a compromise between the optimal freezing rates of the individual organisms. In this situation, the freezing rate is selected to optimize the viability of the various organisms to be frozen together. In certain embodiments, the cryopreservation composition is kept at a temperature of -35°C or lower for a period of at least 24 hours prior to step (b).

[0194] Exemplary cryoprotectants include, for example, acetamide, albumin, ammonium acetate, choline magnesium chloride, sodium bromide, diethyl glycol, dimethylacetamide, dimethylsulfoxide, ethanol, erythritol, glycerol, glucose, formamide, glycerophosphate, proline, pyridine-N-oxide, propylene glycol, ribose, serine, sodium chloride, sodium iodide, sodium sulfate, sorbitol, sucrose, trehalose, triethyene glycol, trimethylamine, valine, xylose. Exemplary cryoprotectants include, for example, alginate, maltodextrin, oligofructose, and the like. It will be appreciated that certain cryoprotectants can also act as lyoprotectants.

[0195] During step (b), the freeze-drying conditions may be selected to optimize the viability of the organism or group of organisms in the cryopreservation mixture.

[0196] In certain embodiments, during step (b), lyophilizing the cryopreservation composition comprises the steps of: (i) equilibrating the cryopreservation composition at a first temperature of about -35°C for about 2 hours; (ii) raising the temperature of the cryopreservation composition to a second temperature of about -25°C to -30°C; (iii) holding the cryopreservation composition at the second temperature and a first pressure of less than or equal to 55 mTorr for about 20 to 72 hours; (iv) raising the temperature of the cryopreservation composition to a third temperature of about 4 to 20°C; and (v) holding the cryopreservation composition at the third temperature and a second pressure of about 100 mTorr for about 20 to 48 hours or until the water activity of the cryopreservation composition is less than about 0.15, thereby producing a cryopreservation composition.

[0197] In certain embodiments, in step (ii), the temperature of the cryopreservation composition is increased to a second temperature of about −25° C. at a heating rate of 0.01° C. / min. In certain embodiments, in step (iv), the temperature of the cryopreservation composition is increased to a third temperature of about 20° C. at a heating rate of about 0.1° C. / min.

[0198] In certain embodiments, during step (b), the cryopreserved composition has a water content of less than about 0.5% (w / w), less than about 1% (w / w), less than about 1.5% (w / w), less than about 2% (w / w), less than about 2.5% (w / w), less than about 3% (w / w), less than about 3.5% (w / w), less than about 4% (w / w), less than about 4.5% (w / w), or less than about 5% (w / w).

[0199] In certain embodiments, during step (b), the cryopreserved composition is from about 1% (w / w) to about 5% (w / w), from about 1.5% (w / w) to about 5% (w / w), from about 2% (w / w) to about 5% (w / w), from about 2.5% (w / w) to about 5% (w / w), from about 3% (w / w) to about 5% (w / w), from about 3.5% (w / w) to about 5% (w / w), from about 4% (w / w) to about 5% (w / w), from about 4.5% (w / w) to about 5% (w / w), from about 1% (w / w), (w / w) to about 4.5% (w / w), about 1% (w / w) to about 4% (w / w), about 1% (w / w) to about 3.5% (w / w), about 1% (w / w) to about 3% (w / w), about 1% (w / w) to about 2.5% (w / w), about 1% (w / w) to about 2% (w / w), about 1% (w / w) to about 1.5% (w / w), about 1.5% (w / w) to about 4.5% (w / w), about 1.5% (w / w) to about 4% (w / w), about 1.5% (w / w) to about 3.5% (w / w), about 1.5% (w / w) to about 3% (w / w), about 1.5% (w / w) to about 2.5% (w / w), about 1.5% (w / w) to about 2% (w / w), about 2% (w / w) to about 4.5% (w / w), about 2% (w / w) to about 4% (w / w), about 2% (w / w) to about 3.5% (w / w), about 2% (w / w) to about 3% (w / w), about 2% (w / w) to about 2.5% (w / w), about 2.5% (w / w) to about 4.5% (w / w), about 2.5% ( In certain embodiments, during step (b), the cryopreserved composition has a water content in the range of about 2% (w / w) to about 4% (w / w), about 2.5% (w / w) to about 3.5% (w / w), about 2.5% (w / w) to about 3% (w / w), about 3% (w / w) to about 4.5% (w / w), about 3% (w / w) to about 4% (w / w), about 3% (w / w) to about 3.5% (w / w), about 3.5% (w / w) to about 4.5% (w / w), about 3.5% (w / w) to about 4% (w / w), or about 4% (w / w) to about 4.5% (w / w). In certain embodiments, during step (b), the cryopreserved composition has a water content in the range of about 2% (w / w) to about 4% (w / w) or about 2% (w / w) to about 3% (w / w). In certain embodiments, the composition has a moisture content in the range of about 1.5% (w / w) to about 4% (w / w).

[0200] In certain embodiments, during step (b), the cryopreserved composition has a water content of about 0.5% (w / w), about 1% (w / w), about 1.5% (w / w), about 2% (w / w), about 2.5% (w / w), about 3% (w / w), about 3.5% (w / w), about 4% (w / w), about 4.5% (w / w), or about 5% (w / w).

[0201] In various embodiments, the method further comprises packaging the cryopreserved composition in a capsule, tablet, or caplet, as described above, which may be coated with a delayed release coating.

[0202] In various embodiments, the method optionally further comprises various steps including: (c) sieving the cryopreserved composition through a #20 US mesh (0.85 mm) screen; (d) mixing the cryopreserved composition; and (e) packaging the cryopreserved composition into a capsule, tablet, or caplet.

[0203] In various embodiments, the method optionally further comprises the steps of (c) sieving the cryopreserved composition through a #20 US mesh (0.85 mm) screen; (d) mixing the cryopreserved composition with one or more pharma- ceutically acceptable excipients to produce a pharmaceutical composition described herein; and (e) packaging the pharmaceutical composition into a capsule, tablet, or caplet.

[0204] 5. Methods of Use and Treatment The compositions described herein can be used in a method of adjusting the gastrointestinal microbiome of a subject in need thereof.The method generally comprises administering to the subject an effective amount of a preserved composition described herein, for example a cryopreserved composition, for example in the form of a pharmaceutical composition or a food product / probiotic.Depending on the particular situation, the method comprises administering to the subject an effective amount of a pharmaceutical composition described herein, or the method comprises administering to the subject an effective amount of a food product or probiotic described herein.

[0205] The preserved (e.g., cryopreserved) compositions described herein can be used to treat a variety of conditions, diseases, or disorders, such as a gastrointestinal condition, disease, or disorder, which can be a small intestinal condition, disease, or disorder, such as Crohn's disease or ulcerative colitis. EXAMPLES

[0206] In order that the disclosure described herein may be more fully understood, the following examples are set forth. The process and analytical examples described in this application are provided to illustrate the compositions and methods provided herein and should not be construed as limiting the scope thereof in any manner. EXAMPLES

[0207] Assay protocol Viable cell count / Total cell count / Non-viable cell count: The assay used herein uses Molecular Probe's LIVE / DEAD® BacLight™ Bacterial Viability Kit, a dual-color fluorescent assay of bacterial viability. This kit uses a mixture of SYTO® 9 green fluorescent nucleic acid stain and a red fluorescent nucleic acid stain, propidium iodide. The stains differ both in their spectral characteristics and in their ability to penetrate healthy bacterial cells. When used alone, SYTO® 9 stain generally labels all bacteria in a population, i.e., those with intact membranes and those with damaged membranes. In contrast, propidium iodide only penetrates bacteria with damaged membranes, causing a decrease in SYTO® 9 stain fluorescence when both dyes are present. Thus, with the appropriate mixture of SYTO® 9 and propidium iodide stain, bacteria with intact cell membranes are stained fluorescent green, while bacteria with damaged membranes are stained fluorescent red. The excitation / emission maxima of these dyes are approximately 480 / 500 nm for SYTO® 9 staining and 490 / 635 nm for propidium iodide. The background remains virtually non-fluorescent. Fluorescence intensity is measured using, for example, a Synergy HT multidetection microplate reader (BioTek®) with an excitation filter of 488 / 20 nm (for both SYTO® 9 and propidium iodide) and emission filters of 528 / 20 nm (emission wavelength of SYTO9) and 645 / 40 nm (emission wavelength of propidium iodide).

[0208] To perform the method, equal volumes of component A (SYTO® 9 dye, 3.34 mM) and component B (propidium iodide, 20 mM) of the kit were combined and mixed thoroughly, after which 3 μL of dye mixture was added per 1 mL of bacterial suspension (8-10 mg / mL). The dye mixture and bacterial suspension were then mixed thoroughly and incubated at room temperature in the dark for 15 minutes. 5 μL of the stained bacterial suspension was then spread onto a microscope slide with a cover slip and observed using a fluorescent microscope with filters set according to the manufacturer's instructions.

[0209] The appearance of the organism is as specified in the United States Pharmacopeia (USP) <631> (Color and Achromicity).

[0210] water content Total water (free and bound) in the lyophilized powder was determined using the Karl Fischer method according to USP <921> (Water Determination).

[0211] water activity The water activity of the cryopreserved powders described below was determined using a method based on the AOAC International Official Method 978.18 (AOAC International Official Methods of Analysis, 17th Edition, AOAC International) in accordance with USP <1112> The tests were performed according to the guidance given in Application of Water Activity Determination to Nonsterile Pharmaceutical Products.

[0212] Total aerobic microbial count USP <61> According to (Microbiological Examination of Nonsterile Products: Microbial Enumeration Tests), a Total Aerobic Microbial Count (TAMC) was used to determine the total number of aerobic bacteria present in rehydrated frozen samples at mesophilic temperatures (30-37°C).

[0213] Combined yeast and mold counts The total yeast and mold count combined (TYMC) is <62> The samples were tested according to (Microbiological Examination of Nonsterile Products: Tests for Specified Microorganisms).

[0214] Optical density (OD at 600 nm) of frozen samples Using a 1 cc syringe and a 21 gauge needle, 1 mL of cryopreserved sample was taken from the growing culture vial / bottle. Prior to sampling, the contents of the vial were mixed thoroughly and the syringe was inserted into a CO 2 After that, 1 mL of the sample was dispensed into a microcentrifuge tube and the optical density was measured at 600 nm (OD 600 ) was recorded. 600 If the OD was less than 0.5, the cell suspension was directly dispensed into a cuvette and the optical density was measured at 600 nm. 600 If was higher than 0.5, OD 600 Distill H until H is less than 0.5 2 Serial dilutions were prepared in O and measurements were recorded.

[0215] pH of frozen samples The pH of the frozen samples was determined according to USP <791> (The standardized test method for pH measurements used in the pharmaceutical, food and beverage, patient care, and dietary supplement industries).

[0216] Osmolality of frozen samples The osmolality of the frozen samples was determined according to USP <785> (osmolality and osmolarity).

[0217] Viscosity of frozen samples The viscosity of the frozen samples was measured according to USP <911> (viscosity) was used to test.

[0218] Thermal characterization of cryopreserved samples (i) Differential Scanning Calorimetry (DSC) Method The DSC instrument used for thermal characterization (e.g., determination of glass transition temperature (Tg')) of the cryopreserved samples described in the Examples herein was a TA Instruments Discovery DSC25 modulated DSC equipped with a refrigerated cooling system.

[0219] Prior to characterization of the cryopreserved samples, the calibration of the DSC instrument was checked with a 1% sodium chloride solution. The sodium chloride calibration sample was determined to have an onset of melting of -21.71°C (theoretical = -21.5°C).

[0220] In preparation for freezing for DSC analysis, a pipette was used to pipet an 18 μL aliquot of the liquid to be analyzed (eg, sodium chloride calibration samples or liquid samples of cryopreservation solution) into an aluminum DSC sample pan and sealed.

[0221] DSC Method: Sample and reference pans (empty aluminum pans) were loaded into the DSC at 40° C. and then cooled at 10° C. / min to −90° C. The pans were then held at −90° C. for 2 minutes and subsequently heated to 30° C. at 10° C. / min.

[0222] (ii) Freeze-drying microscopy The freeze-drying microscopy equipment included a Nikon Eclipse LV100Npol polarizing microscope equipped with a Linkam FDCS 196 thermal stage, a T96 controller, an LNP96 liquid nitrogen pump, and an Edwards E2M1.5 vacuum pump.

[0223] Method: A 5 μL aliquot of sample (e.g. liquid sample of cryopreservation solution) was placed on a glass slide placed on a thermal stage after applying a small drop of silicon oil. A cover glass was placed on top of the sample and the chamber was sealed. The sample was cooled to -45°C at 10°C / min, held for 2 minutes and a micrograph was taken. The vacuum was started and the sample was held at a fixed temperature of -45°C until the sublimation surface was visible. Then another micrograph was taken. The sample was then warmed to -10°C at 10°C / min. Micrographs were collected as the sample went through various thermal transitions (e.g. collapse temperature (Tc)). EXAMPLES

[0224] Cell culture and harvesting This example describes the procedures used to culture and harvest each of the organisms described herein.

[0225] The cell bank was thawed in a room temperature water bath for approximately 15 minutes. A filtered blunt needle was prepared with anaerobic gas flowing into a 50 mL Falcon tube. The syringe was then filled with anaerobic gas (e.g., N 2 , CO 2 The plate was flushed with 500 mL of argon (argon) to remove oxygen. Using a needle, the amount of thawed bank medium required for the desired percent inoculum was introduced into a vial or bottle of anaerobic medium.

[0226] Media components were weighed, water was added to the specified volume, mixed until dissolved, and then autoclaved at 121°C using a liquid cycle for 60 minutes or sterile filtered into sterile containers. Media was stored in 100% CO 2 or N 2The medium was sparged with anaerobic gas at 2 L / min with an agitation speed of 150 rpm (6-blade Rushton impeller). The set point was set at 37° C. and sparging was continued at 2 L / min until the redox potential was sufficiently negative, i.e., in the range of about −50 mV to about −500 mV. After the redox potential met the specified criteria, the gas was switched from sparging the medium to purging the headspace with anaerobic gas at 1 L / min, and then an overflow of 1 L / min was maintained.

[0227] 2L anaerobic bottles were placed at the specified OD 600 When the initial pH reached 10, the culture was ready to be transferred to the fermenter. 100 mL of culture medium was then removed from the bottle and added to the fermenter through the inoculation port. Fermentation was allowed to continue for the specified period of time.

[0228] Harvesting was performed through a Repligen Hollow Fiber PES 750 kDa membrane and the 10 L was concentrated to approximately 400-600 mL. The retentate was collected and filled into centrifuge bottles in an anaerobic chamber. The retentate was centrifuged at 10,000 rpm for 30 minutes and the cell paste was collected in the anaerobic chamber for further processing. EXAMPLES

[0229] Preparation of a preservation composition for anaerobic microorganisms For each excipient blend listed in Table 1 below, all dry components (e.g., raftilose, maltodextrin, sodium alginate, and trehalose and / or sucrose) were dry mixed until the powder mixture appeared homogenous. The powder mixture was then slowly added to deionized (DI) water in a second container and stirred until all components were dissolved. The resulting solution was then placed in an anaerobic chamber and stirred continuously for 60 minutes. The pH of the excipient blend was adjusted using either NaOH or HCl, while monitoring the pH using a pH meter, until the pH reached an optimal level for the organism / organisms. The excipient blend was then sterile filtered into a sterile container using a 0.45 micron filter.

[0230] In the anaerobic chamber, frozen cell paste (cell concentrate) or fresh cell paste was placed in a sterile container. The cell paste was mixed with anaerobic buffer, spent medium, or fresh medium in a 1:1, 1:2, or 1:3 ratio, as desired. The mixture was mixed for 20-30 minutes, and the resulting blend was poured into lyophilization trays to a defined filling depth. The lyophilization trays were placed into plastic bags and laminated foil pouches and finally heat sealed, all in the anaerobic chamber to maintain a reduced redox potential within the range of -50 to -500 mV. The sealed trays were then placed into a freezer or lyophilization shelf to freeze and were left frozen for at least 24 hours before starting the lyophilization.

[0231] [Table 1] EXAMPLES

[0232] Preparation and Efficacy Testing of Compositions for Cryopreserving Anaerobic Microorganisms The cryopreserved composition prepared as in Example 3 was then lyophilized as follows:

[0233] Prior to lyophilization, the frozen compositions were frozen at temperatures between -35°C and -150°C and then placed in a freeze dryer (Virtis, Genesis SQ Super XL70) and equilibrated at -35°C. A three-step gradient cycle was used to freeze-dry the frozen compositions, and the freeze-drying process was terminated when the water activity was <0.15 during secondary drying. The freeze-drying parameters are listed in Table 2, and the temperature / pressure profile used during the freeze-drying cycle is shown in Figure 1. The freeze-drying cycle lasted approximately 90 hours in total. The freeze-dried cakes were then stored at -20°C until further processing.

[0234] The glass transition temperature, Tg', of the freeze-dried cake was determined to be -24°C.

[0235] [Table 2]

[0236] The lyophilized cake was then sieved through a #20 US mesh (0.85 mm) screen followed by mixing in the sieve pan for 2 minutes. All sieving, mixing and packaging was carried out in a nitrogen glove box or anaerobic chamber in an atmosphere with a relative humidity (RH) of ≦15%. The resulting material (drug substance) was dispensed and packaged in polyethylene bags and placed in a foil laminate pouch along with a drying pouch and then sealed. The sealed pouch with the drug substance was stored below −20° C. Samples were drawn and tested for % viability.

[0237] Viability results for the three cryopreservation mixtures (ACP-001, ACP-002, and ACP-003) compared to two commercially available blend mixtures (CP-001 and CP-002) (OPS DIagnostics catalog numbers LR2X-500-02 and MFDB-500-06) are presented in Table 3.

[0238] [Table 3] JPEG2024515796000005.jpg230166

[0239] The results from Table 3 show that for the various bacterial families and genera tested, the efficacy (viability) produced for the three ACP blends was better than that achieved using two commercially available blends (OPS DIagnostics catalog numbers LR2X-500-02 and MFDB-500-06). EXAMPLES

[0240] Stability of cryopreserved compositions Freeze-thaw stability Each freeze-thaw cycle was performed by removing the sample from storage at -20±5°C in its packaging and placing it at ambient room temperature (22°C) for at least 6 hours. After thawing, the samples were returned to storage at -20±5°C for 12-24 hours. To ensure that all samples were analyzed simultaneously, samples were staggered through up to four freeze-thaw cycles.

[0241] The results are summarized in Table 4.

[0242] [Table 4]

[0243] The results of this study indicate that the drug substance (for all four bacterial families tested) is stable for at least four freeze-thaw cycles from the recommended storage temperature (-20 ± 5°C) to room temperature (22°C).

[0244] Freezing cycle stability Each temperature cycle was performed by removing the sample in its packaging from storage at -20±5°C and placing it on dry ice for 2 hours. After equilibration at -80°C, the samples were returned to storage at -20±5°C for 2 hours. To ensure that all samples were analyzed simultaneously, samples were alternately put through up to four freeze-thaw cycles.

[0245] The results are summarized in Table 5.

[0246] [Table 5]

[0247] The results of this study show that the drug substance (for all four bacterial families tested) was stable upon low temperature exposure at -80°C and at least four temperature cycles between the recommended storage temperature (-20 ± 5°C) and lower temperatures (-80°C).

[0248] long term stability Long-term stability studies of the cryopreserved compositions prepared in Example 4 were performed for each storage condition (control vs. accelerated) using the analytical methods provided in Table 6.

[0249] The stability study was performed according to ICH Q1 and ICH Q5 guidelines.

[0250] [Table 6]

[0251] For clarity, only potency data (CFU / g of drug substance powder) are presented for material stored at 2-8°C and 25°C. Additionally, batches of material that had less than 5% or more than 5% total moisture were included in the stability program to determine the effect of moisture content on stability.

[0252] The results are shown in Tables 7 to 9.

[0253] Table 7 shows the stability (Log scale of CFU / g) at 2-8° C. where the bulk powder has a total moisture content of less than 5% (w / w).

[0254] [Table 7]

[0255] Table 8 shows the stability (Log scale of CFU / g) at 25° C. where the bulk drug powder has a total moisture content of less than 5% (w / w).

[0256] [Table 8]

[0257] Table 9 shows the stability (Log scale of CFU / g) at 25° C. for bulk drug powders having a total moisture content greater than 5% but less than 8%.

[0258] [Table 9]

[0259] The results in Tables 7-9 indicate that maintaining a moisture content below 5% is important for long-term stability. All organisms tested stored at or below 5% (w / w) moisture content were stable for 24 months when stored at both 2-8°C and 25°C. EXAMPLES

[0260] Characterization of cryopreservation compositions Air exposure of API Evaluation of the oxygen sensitivity of the cryopreserved compositions prepared in Example 4 was performed by subjecting samples in sealed polyethylene bags (sealed in ambient air) to a temperature of 22° C. and a relative humidity of 19% via the use of a saturated 40% NaOH solution for up to 7 days. Sample exposure was staggered across time points to ensure that all samples were analyzed simultaneously.

[0261] Appearance, moisture content, and viability tests of the drug substance were measured as a function of time in the presence of air. The results of this study are summarized in Table 10.

[0262] [Table 10]

[0263] The results of this study show that for all four bacterial families tested, the drug substance was stable when stored for up to seven days at 22°C and 19% relative humidity in the presence of atmospheric oxygen levels (approximately 20.95%). When stored in a dry, dormant state, the organisms are less sensitive to oxygen exposure.

[0264] Drug substance moisture adsorption research The characteristics and effects of moisture uptake were evaluated under the conditions described in Table 11. Relative humidity conditions were maintained through the use of saturated salt solutions and confirmed using an Acurite digital temperature / hygrometer to which the various samples were exposed.

[0265] [Table 11]

[0266] Samples were exposed to the conditions for periods of 2, 4, 8, and 24 hours in addition to an initial test prior to exposure.

[0267] The results are summarized in Tables 12-15 and the organisms were tested for appearance, moisture content, and viability.

[0268] [Table 12]

[0269] [Table 13]

[0270] [Table 14]

[0271] [Table 15]

[0272] The results described in this example demonstrate that exposure to increasing humidity levels results in increasing amounts of moisture in the freeze-dried material. Moisture absorption is most evident at higher humidity levels and longer exposure times.

[0273] The results indicate that, associated with increasing moisture content of the freeze-dried material, there appears to be a corresponding loss in viability with increasing humidity and duration of exposure. Control samples, both at the higher temperature (22° C.) and at the recommended storage conditions (−20±5° C.), showed no increase in moisture content or loss in viability, suggesting that the impact on product quality is a result of moisture content.

[0274] Furthermore, a relationship appears to exist between exposure to elevated humidity and loss of viability of organisms. Loss of viability is accompanied by a corresponding shift from a free-flowing powder to a cake-like appearance, consistent with moisture absorption. Although increases in moisture value correlate with loss of viability, the changes in absolute moisture content (%wt / wt) are actually small, suggesting that initial moisture level is a critical quality attribute and that the processing environment is a critical process parameter that requires control.

[0275] Incorporated by reference This application refers to various issued patents, published patent applications, academic papers, and other publications, all of which are incorporated herein by reference. Moreover, any particular embodiment of the present disclosure that is within the scope of the prior art may be expressly excluded from any one or more of the claims. Such embodiments may be excluded even if the exclusion is not expressly stated herein, since they are deemed known to those skilled in the art. Any particular embodiment of the present disclosure may be excluded from any claim for any reason, regardless of whether it is related to the existence of prior art.

[0276] Equivalent The present invention may be embodied in other specific forms without departing from its spirit or essential characteristics. The foregoing embodiments are therefore to be considered in all respects as illustrative rather than limiting of the invention described herein. The scope of the invention is therefore indicated by the appended claims, rather than by the foregoing description, and all changes that come within the meaning and range of equivalency of the claims are intended to be embraced therein.

Claims

1. 1. A composition for preserving anaerobic microorganisms comprising a viscosity modifier and an energy source for the microorganisms, the composition having, when solvated in an aqueous solvent, a viscosity in the range of about 5 cP to about 6,000 cP, an osmolality in the range of about 300 Osm / L to about 4,000 Osm / L, a pH in the range of about 4 to about 8, and optionally an oxidation-reduction potential in the range of -50 mV to -500 mV.

2. The composition of claim 1 , wherein the composition is a cryopreservative composition.

3. 1. A cryopreservation composition for cryopreserving anaerobic microorganisms comprising a viscosity modifier and an energy source for the microorganisms, the cryopreservation composition having, when solvated in an aqueous solvent, a viscosity in the range of about 5 cP to about 6,000 cP, an osmolality in the range of about 300 Osm / L to about 4,000 Osm / L, a pH in the range of about 4 to about 8, and optionally an oxidation-reduction potential in the range of -50 mV to -500 mV.

4. 4. The composition of claim 1, further comprising one or more anaerobic microorganisms.

5. 4. The composition of any one of claims 1 to 3, wherein the viscosity is in the range of about 50 cP to about 4,000 cP, about 100 cP to about 2,000 cP, or about 100 cP to about 1,000 cP.

6. 4. The composition of claim 1, wherein the osmolality is in the range of about 300 Osm / L to about 3,000 Osm / L, about 400 Osm / L to about 3,000 Osm / L, or about 500 Osm / L to about 3,000 Osm / L.

7. 4. The composition of claim 1, wherein the pH is in the range of about 4 to about 8, about 5 to about 8, about 4 to about 7.5, about 5 to about 7.5, or about 5.5 to about 7.

5.

8. 4. The composition of claim 1, wherein the composition comprises from about 0.01% (w / w) to about 0.4% (w / w) of the viscosity modifier when solvated in an aqueous solvent.

9. The composition of claim 1 , wherein the viscosity modifier is a polymeric composition.

10. 10. The composition of claim 9, wherein the polymer composition comprises alginate, acacia, alginic acid, agar, aramic acid, bentonite, carbomer, carrageenan, cellulose, dextrin, gelatin, gellan gum, guar gum, carboxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, methyl cellulose, mucin, pectin, starch, xanthan gum, or combinations thereof.

11. The composition of claim 10 , wherein the polymer composition comprises an alginate.

12. The composition of claim 11 , wherein the alginate is sodium alginate.

13. 13. The composition of claim 12, wherein the sodium alginate has a viscosity range of about 250 cP to about 14,000 cP.

14. 4. The composition of claim 1, wherein the energy source is a cell culture medium that allows the growth of the microorganism.

15. 4. The composition of any one of claims 1 to 3, wherein the composition comprises from about 3% (w / w) to about 15% (w / w) of the energy source when solvated in an aqueous solvent.

16. 4. The composition of claim 1, wherein the energy source is a monosaccharide, a disaccharide, an oligosaccharide, or a combination thereof.

17. 17. The composition of claim 16, wherein the monosaccharide is selected from the group consisting of glucose, fructose, xylose, and combinations thereof.

18. 17. The composition of claim 16, wherein the disaccharide is trehalose, sucrose, or a combination thereof.

19. The composition of claim 16 , wherein the oligosaccharide is an inulin derivative.

20. The composition of claim 17, wherein the inulin derivative is a fructooligosaccharide.

21. The composition of claim 1 , further comprising a protein stabilizer.

22. 22. The composition of claim 21, wherein the composition comprises from about 5% (w / w) to about 10% (w / w) of the protein stabilizing agent when solvated in an aqueous solvent.

23. 22. The composition of claim 21, wherein the protein stabilizing agent is selected from the group consisting of amino acids, antioxidants, disaccharides, reducing agents, polyethylene glycol, and combinations thereof.

24. 24. The composition of claim 23, wherein the disaccharide is selected from the group consisting of trehalose, sucrose, isomaltose, lactose, maltose, lactulose, cellobiose, and combinations thereof.

25. 5. The composition of claim 4, wherein the one or more anaerobic microorganisms are present in the form of a cell paste or a cell slurry.

26. 26. The composition of claim 25, wherein the composition comprises from about 30% (w / w) to about 50% (w / w) of the cell paste or cell slurry when solvated in an aqueous solvent.

27. (i) a cell paste comprising about 30% (w / w) to about 50% (w / w) of one or more anaerobic microorganisms; (ii) about 3% (w / w) to about 15% (w / w) of an energy source for the microorganism; (iii) from about 0.01% (w / w) to about 0.4% (w / w) of a viscosity modifier; and (iv) about 30% (w / w) to about 60% (w / w) of an aqueous solvent; and having a viscosity in the range of about 5 cP to about 6,000 cP, an osmolality in the range of about 300 Osm / L to about 4,000 Osm / L, and a pH in the range of about 4 to about 8.

28. (i) a cell paste comprising about 30% (w / w) to about 50% (w / w) of one or more anaerobic microorganisms; (ii) about 0.5% (w / w) to about 5% (w / w) of raftilose; (iii) about 3% (w / w) to about 10% (w / w) of maltodextrin; (iv) about 0.01% (w / w) to about 0.4% (w / w) sodium alginate; (v) about 5% (w / w) to about 10% (w / w) trehalose; and (vi) 0% (w / w) to about 5% (w / w) sucrose; and (vii) about 35% (w / w) to about 55% (w / w) water; and having a viscosity in the range of about 5 cP to about 6,000 cP, an osmolality in the range of about 300 Osm / L to about 4,000 Osm / L, and a pH in the range of about 4 to about 8.

29. 29. The composition of claim 27 or 28, wherein the composition has a redox potential in the range of -50 mV to -500 mV.

30. 30. The composition of any one of claims 1 to 3, 27 and 28, further comprising a buffering agent.

31. A preserved composition comprising one or more anaerobic microorganisms, a viscosity modifier, an energy source for said microorganisms, an oxidation-reduction potential in the range of -50 mV to -500 mV, and a moisture content of less than 5% (w / w).

32. 32. The composition of claim 31, wherein the composition is a cryopreserved composition.

33. 1. A cryopreservation composition comprising one or more anaerobic microorganisms, a viscosity modifier, an energy source for said microorganisms, an oxidation-reduction potential in the range of -50 mV to -500 mV, and a water content of less than 5% (w / w).

34. 34. The composition of any one of claims 31 to 33, wherein the moisture content is in the range of about 2% (w / w) to about 4% (w / w) or about 2% (w / w) to about 3% (w / w).

35. 34. The composition of any one of claims 31 to 33, wherein the viscosity modifier is a polymeric composition.

36. 36. The composition of claim 35, wherein the polymer composition comprises alginate, acacia, alginic acid, agar, aramic acid, bentonite, carbomer, carrageenan, cellulose, dextrin, gelatin, gellan gum, guar gum, carboxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, methyl cellulose, mucin, pectin, starch, xanthan gum, or a combination thereof.

37. The composition of claim 35 , wherein the polymer composition comprises an alginate.

38. 37. The composition of claim 36, wherein the alginate is sodium alginate.

39. 39. The composition of claim 38, wherein the sodium alginate has a viscosity range of 250 to 14,000 cP.

40. 34. The composition of any one of claims 31 to 33, wherein the energy source is a cell culture medium that allows growth of the microorganism.

41. 34. The composition of any one of claims 31 to 33, wherein the energy source is a monosaccharide, a disaccharide, an oligosaccharide, or a combination thereof.

42. 42. The composition of claim 41, wherein the monosaccharide is selected from the group consisting of glucose, fructose, xylose, and combinations thereof.

43. 42. The composition of claim 41, wherein the disaccharide is selected from the group consisting of trehalose, isomaltose, lactose, maltose, lactulose, cellobiose, sucrose, and combinations thereof.

44. 42. The composition of claim 41, wherein the oligosaccharide is an inulin derivative.

45. 45. The composition of claim 44, wherein the inulin derivative is a fructooligosaccharide.

46. 34. The composition of any one of claims 27, 28, and 31-33, further comprising a protein stabilizer.

47. 47. The composition of claim 46, wherein the protein stabilizing agent is selected from the group consisting of amino acids, antioxidants, disaccharides, reducing agents, polyethylene glycol, and combinations thereof.

48. 48. The composition of claim 47, wherein the disaccharide is selected from the group consisting of trehalose, sucrose, isomaltose, lactose, maltose, lactulose, cellobiose, and combinations thereof.

49. 34. The composition of any one of claims 31 to 33, further comprising a buffering agent.

50. (i) one or more anaerobic microorganisms; (ii) an energy source comprising raftilose and maltodextrin; (iii) sodium alginate; (iv) a protein stabilizing agent comprising trehalose, sucrose, and combinations thereof; (vi) a moisture content of less than 5% (w / w); A cryopreserved composition comprising:

51. 51. The composition of claim 50, wherein the composition has a redox potential in the range of -50 mV to -500 mV.

52. The one or more anaerobic microorganisms are selected from the group consisting of Akkermansiaceae, Bacillaceae, Bacteroidetes, Bifidobacteriaceae, Christensenellaceae, Clostridiaceae, Coriobacteriaceae, Eggerthellaceae, Enterobacteriaceae, Eubacteriaceae, Lachnospira 52. The composition of any one of claims 1 to 3, 31 to 33, and 50 to 51, wherein the bacterial strain is selected from the group consisting of Lactobacillaceae, Lactobacillaceae, Leuconostocaceae, Odoribacteraceae, Oxalobacteraceae, Porphyromonadaceae, Prevotellaceae, Rikenellaceae, Ruminococceae, Streptococceae, and combinations thereof.

53. 52. The composition of any of claims 31-33 and 50-51, wherein when stored at 4°C for 6 or 12 months and then rehydrated, the composition results in less than 3, 2, 1, or 0.5 log loss of colony forming units (CFU).

54. 52. The composition of any one of claims 31-33 and 50-51, wherein the composition remains substantially stable at 4°C for about 2 years.

55. 52. The composition of any one of claims 31 to 33 and 50 to 51, wherein when rehydrated in an aqueous fluid, the composition allows regrowth of the one or more microorganisms within 5, 4, 3, or 2 hours of rehydration.

56. 52. A pharmaceutical composition comprising the cryopreserved composition of any one of claims 31-33 and 50-51, optionally disposed within a capsule, tablet, or caplet.

57. 52. A food product or probiotic comprising the cryopreserved composition of any one of claims 31-33 and 50-51, optionally disposed within a capsule, tablet, or caplet.

58. 52. A method of modulating the gastrointestinal microbiome in a subject in need thereof, comprising administering to the subject an effective amount of a composition described in any one of claims 31-33 and 50-51.

59. (a) combining an anaerobic microorganism with an excipient blend comprising an aqueous mixture comprising a viscosity modifier and an energy source for said microorganism to produce a cryopreservation mixture, said cryopreservation mixture having a viscosity in the range of about 5 cP to about 6,000 cP, an osmolality in the range of about 300 Osm / L to about 4,000 Osm / L, a pH in the range of about 4 to about 8, and optionally an oxidation-reduction potential in the range of −50 mV to −500 mV; (b) lyophilizing the cryopreservation mixture to reduce the water content of the cryopreservation mixture to less than about 5% (w / w) to produce the cryopreservation composition.

52. A method of producing a cryopreserved composition according to any one of claims 31 to 33 and 50 to 51, comprising:

60. 60. The method of claim 59, wherein during step (a), the microorganism is in the form of a cell paste.

61. 60. The method of claim 59, wherein the cell paste comprises the microorganism in cell culture medium.

62. 60. The method of claim 59, further comprising packaging the cryopreserved composition in a capsule, tablet, or caplet.

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