Ferritin compositions and uses thereof

Abstract

The present disclosure relates to the use of ferritin in lieu of transferrin in culture medium and other compositions or products, such as biomass compositions or cultivated meat compositions. Such compositions or products are suitable for human and / or pet consumption. The invented culture medium is not only able to support long-term growth and culture of animal cells ex vivo but, unlike transferrin-containing culture media, is affordable for both pharmaceutical and food production. The present disclosure also encompasses methods and processes in making and / or manufacturing such cell culture media and / or compositions, as well as their use in products for human and / or pet consumption. The products include for example, food products, feed products, nutraceutical products, and pharmaceutical products.

Description

Attorney Docket No.: 116673-856389FERRITIN COMPOSITIONS AND USES THEREOFCross Reference to Related Application

[0001] The present application claims the benefit of U.S. Provisional Patent Application No. 63 / 690,241 , entitled, “FERRITIN COMPOSITIONS AND USES THEREOF” filed September 3, 2024. The contents of the aforementioned application is hereby incorporated by reference in its entirety.Field of the Invention

[0002] The present disclosure generally relates to cell culture and compositions or products thereof. Such compositions or products are usable in food, dietary supplements, and / or pharmaceutical industries for human and pet consumption.Background of the Invention

[0003] Growing cells ex vivo requires a culture medium that is able to supply essential elements for cell growth, such as a carbon-based energy source (e.g. glucose), amino acids, salts, vitamins, minerals and water. Such culture medium needs to support at least cell viability and proliferation, with most formulations being cost prohibitive for many applications. There is a need to develop functional culture media that are cost efficient and suitable for mass production.Summary of the Invention

[0004] One aspect of the present disclosure encompasses a cell biomass composition, and the composition is produced ex vivo by culturing a cell line in a culture medium comprising ferritin and substantially free of transferrin. In one aspect, the ferritin is a food-grade, feed-grade, or pharmaceutical grade. In another aspect, the ferritin comprises an animal-derived ferritin, a recombinant ferritin, a plant-derived ferritin, or any combination thereof. In another aspect, the plant comprises a legume plant, and preferably being a Glycine max or Pisum sativum. In one aspect, the ferritin is in an amount from about 50 pg / ml to about 150 pg / ml. In another aspect, the ferritin is in an amount from about 80 pg / ml to about 120 pg / ml. In another aspect, ferritin is1105616786.2Attorney Docket No.: 116673-856389 in an amount of about 100 pg / ml. In yet another aspect, composition is suitable for human or pet consumption. In one aspect, the composition is free of any animal ingredient or is plant based. In yet another aspect, the composition comprises animal ingredients. In one aspect, the composition is serum-free or serum-containing. In one aspect, the composition is incorporated into a food product, a feed product, a nutraceutical product, a dietary supplement product, a performance-enhancing product, a veterinary product, a chemical product, a scientific reagent product or a pharmaceutical product. In another aspect, the composition is incorporated in a cultivated meat product. In one aspect, the cultivated meat product further comprises a functional ingredient to supplement the product’s protein content, lipid content, sugar content, fiber content, calcium content, iron content, or any combination thereof. In yet another aspect, the cultivated meat product further comprises a food grade or feed grade flavorant, a food grade or feed grade colorant, a food grade or feed grade preservative, a food grade or feed grade antioxidant, or any combination thereof. In another aspect, the food grade or feed grade flavorant comprises a chicken flavorant, a beef flavorant, a pork flavorant, a fish flavorant, a smoke meat flavorant, a mouse flavorant, or any combination thereof. In another aspect, the composition is lab grade, food grade, feed grade, pharmaceutical grade, or chemical grade. In another aspect, the composition is used as a culture medium component, as a carrier component, as an excipient component, as a reagent, or any combination thereof. In another aspect, the composition is used as a culture medium for culturing and / or proliferating cells suitable for a cultivated meat product. In yet another aspect, the cells suitable for a cultivated meat product comprise stem cells, fibroblast cells, immortalized cells, or any combination thereof.

[0005] Another aspect of the present disclosure encompasses a cultivated meat composition comprising ferritin and substantially free of transferrin. In one aspect, the composition comprises (i) ferritin; (ii) a culture medium substantially free of transferrin; and (iii) cells, wherein the cells are capable of being cultured and / or proliferated in the culture medium and ferritin. In one aspect, the ferritin is in an amount sufficient to support the cell line growth. In another aspect, the ferritin is in an amount from about 50 pg / ml to about 150 pg / ml. In yet another aspect, the ferritin is in an amount from2105616786.2Attorney Docket No.: 116673-856389 about 80 pg / ml to about 120 pg / ml. In yet another aspect, the ferritin is in an amount of about 100 pg / ml. In one aspect, the ferritin comprises an animal-derived ferritin, a recombinant ferritin, a plant-derived ferritin, or any combination thereof. And the plant comprises a legume plant, and preferably being a Glycine max or Pisum sativum. In one aspect, the cell line comprises a stem cell, a fibroblast cell, an immortalized cell line, or any combination thereof. The stem cell comprises an embryonic stem cell line, an induced pluripotent stem cell line, or any combination thereof. And the immortalized cell line comprises a myoblast cell line, an adipocyte cell line, or any combination thereof. In one aspect, the ferritin is suitable for human or pet consumption. In another aspect, the composition is incorporated into a food product, a feed product, a nutraceutical product, a dietary supplement product, a performance-enhancing product, a veterinary product, a pharmaceutical product, or into a reagent to produce a chemical product. In yet another aspect, the cultivated meat composition further comprises a functional ingredient to supplement the composition’s protein content, lipid content, sugar content, fiber content, calcium content, iron content, or any combination thereof. In another aspect, the cultivated meat product further comprises a food grade or feed grade flavorant, a food grade or feed grade colorant, a food grade or feed grade preservative, a food grade or feed grade antioxidant, or any combination thereof. And the food or feed-grade flavorant comprises a chicken flavorant, a beef flavorant, a pork flavorant, a fish flavorant, a smoke meat flavorant, a mouse flavorant or any combination thereof.

[0006] Another aspect of the present disclosure encompasses a method of producing a cultivated meat composition, comprising: (i) obtaining or having obtained a cell line; (ii) culturing the cell line in a transferrin-based culture medium; (iii) weaning the cultured cells into a mixed culture medium comprising the transferrin-based culture medium and a ferritin-based culture medium, wherein the transferrin-based culture medium accounts for about 30% (v / v) to about 50% (v / v) of the mixed culture medium; (iv) passaging the weaned cultured cells of step (iii) into the ferritin-based culture medium; and (v) continuing passaging the cells in the ferritin-based culture medium; (vi) harvesting the cultured cells alone, the cultured cells together with the spent media, or the biomass inside the bioreactor, and thereby producing the cultivated meat3105616786.2Attorney Docket No.: 116673-856389 composition; wherein the transferrin-based culture medium is substantially free of ferritin; and wherein the ferritin-based culture medium is substantially free of transferrin. In one aspect, the mixed medium comprising about 50% (v / v) of the transferrin-based culture medium and about 50% (v / v) of the ferritin-based culture medium. In another aspect, the transferrin-based culture medium is a commercially available culture medium or a non-commercially available culture medium. In one aspect, the transferrin-based culture medium is KnockOut™ Serum Replacement (KSR medium). In another aspect, the ferritin-based culture medium comprises no component from KSR. In another aspect, the ferritin-based medium comprises ferritin in an amount sufficient to support the cell line proliferation. In one aspect, the ferritinbased medium comprises ferritin in an amount from about 50 pg / ml to about 150 pg / ml. In yet another aspect, the ferritin-based medium comprises ferritin in an amount from about 80 pg / ml to about 120 pg / ml. In yet another aspect, the ferritin-based medium comprises ferritin in an amount of about 100 pg / ml. In one aspect, the ferritin in the ferritin-based medium comprises an animal-derived ferritin, a recombinant ferritin, a plant-derived ferritin, or any combination thereof. And the plant comprises a legume plant, and preferably being a Glycine max or Pisum sativum. In one aspect, the ferritin in the ferritin-based medium is suitable for human or pet consumption. In yet another aspect, the cell line comprises a stem cell, a fibroblast cell, an immortalized cell line, or any combination thereof. In yet another aspect, the stem cell comprises an embryonic stem cell, an induced pluripotent stem cell, or any combination thereof. In one aspect, the immortalized cell line comprises a myoblast cell line, an adipocyte cell line, or any combination thereof.

[0007] Another aspect of the present disclosure encompasses cell culture medium comprising ferritin and substantially free of transferrin. In one aspect, the ferritin is in an amount sufficient to support cell proliferation from a cell line. In another aspect, the ferritin is in an amount from about 50 pg / ml to about 150 pg / ml. In one aspect, the ferritin is in an amount from about 80 pg / ml to about 120 pg / ml. In yet another aspect, the ferritin is in an amount of about 100 pg / ml. In another aspect, the ferritin comprises an animal-derived ferritin, a recombinant ferritin, a plant-derived ferritin, or any combination thereof. And the plant comprises a legume plant, and preferably being a4105616786.2Attorney Docket No.: 116673-856389 glycine max or Pisum sativum. In yet another aspect, the cell line comprises a stem cell line, a fibroblast cell line, a cancer cell line, an immortalized cell line or any combination thereof. The stem cell comprises an embryonic stem cell, an induced pluripotent stem cell, or any cell line combination thereof. The cancer cell line comprises a Hela cell line, a CHO cell line, or any cell line combination thereof. The immortalized cell line comprises a myoblast cell line, an adipocyte cell line, or any combination thereof.Brief Description of the Figures

[0008] FIG. 1 shows the relative growth rate of cells in animal-free medium with ferritin, animal-free medium without transferrin or ferritin, and animal-free medium with recombinant transferrin. Growth of cells were determined over passages 47-58. The growth of cells in animal-free medium with recombinant transferrin was set to 100% and used as a baseline for comparison.

[0009] FIG. 2 shows the relative growth rate of cells using the base formulation of the commercially-established animal-free growth media, Essential 8 (E8), where transferrin is replaced with ferritin, where the medium contains neither transferrin nor ferritin, and where the medium contains recombinant transferrin. Growth of cells was determined over passages 57-62. The growth of cells in animal-free E8 medium with recombinant transferrin was set to 100% and used as a baseline for comparison.

[0010] FIGs. 3A-3B illustrate morphology of mouse embryonic stem cell spheroids during long-term culture (images of cells at passage #12) in the cell culture medium free of animal-ingredient, showing equivalent morphology of cell culture with ferritin (and substantially free of transferrin) (FIG. 3A) versus cell culture with transferrin (and substantially free of ferritin) (FIG. 3B).

[0011] FIGs. 4A-4B provides exemplary Approach 1 and 2 for culturing cells in the ferritin-based cell culture medium.

[0012] FIG. 5 is an image of a cat eating a can of Mouse Mousse.Detailed Description5105616786.2Attorney Docket No.: 116673-856389

[0013] The present disclosure is at least partially based on the surprising discovery that ferritin is able to successfully replace transferrin in a cell culture medium, wherein ferritin supports cell viability and proliferation that is equal, if not superior, to that of transferrin. Ferritin is generally considered as an iron storage protein and not an iron delivery protein. Therefore, it was surprising that ferritin can be used as a replacement for transferrin. Due to the significantly lower cost of plant-derived ferritin (currently, plant-derived ferritin priced at around US$0.022 / L to US$0.20 / L) compared to transferrin (currently, transferrin priced at around US$60 / L), this discovery will enable a cell culture medium that is cost effective and, therefore, make it practically feasible for mass production of cell culture and related cultivated meat compositions or biomass compositions. Nonetheless, the ferritin suitable for the current disclosure does not limit to plant-derived or plant-based ferritin, any ferritin from any source is expressly within the scope of the current disclosure, which includes, but not limited to ferritin produced through recombinant technology or ferritin isolated or derived from a non-plant, such as an animal, an alga, a bacterium, or a fungus.I. Ferritin

[0014] The present disclosure relates to ferritin. Ferritin is a universal intracellular protein that stores iron and releases it in a controlled fashion. The protein is produced by almost all living organisms, including archaea, bacteria, algae, higher plants, and animals. It is the primary intracellular iron-storage protein in both prokaryotes and eukaryotes, keeping iron in a soluble and non-toxic form. Ferritin encompassed by the current disclosure include both natural occurring ferritin, and ferritin produced through recombinant technology. In one aspect, ferritin is plant-derived or plant-based ferritin. In another aspect, ferritin is isolated, extracted or derived from a non-plant, such as an animal, a bacterium, an alga, or a fungus. In another aspect, ferritin is produced through recombinant technology. In animals, ferritin is mainly present in tissues, especially in the liver, kidney, spleen and bone marrow erythroid cells where it serves as an iron reserve for the production of hemoglobin. A small fraction of ferritin is in the serum and contributes little to overall iron storage, but is used clinically as a reporter of iron levels in an animal.6105616786.2Attorney Docket No.: 116673-856389

[0015] Ferritin is a major iron storage protein. Intracellular ferritin is composed of24 subunits of L- and H-chains in varied proportions. Extracellular (serum) ferritin is composed mostly, but not exclusively, of L-ferritin. Secretion of L-ferritin has been carefully documented in cell cultures. Structure wise, ferritin is a hollow globular protein of mass 474 kDa and comprising 24 subunits. Typically, it has internal and external diameters of about 8 and 12 nm, respectively. The nature of these subunits varies by class of organism. Ferritin protein subunits, four a-helix bundles, contain a catalytic center that converts two Fe(ll) atoms to an Fe(lll)-oxo bridged dimer intermediate in mineralization. The two classes of ferritins are (i) maxi-ferritins, 24- polypeptide, 4-bundle subunit assemblies found in animals, plants, and bacteria; and (ii) mini-ferritins (also called Dps proteins), 12-polypeptide, 4-bundle subunit assemblies in archaea and bacteria. Ferritin is an intracellular, protein-iron complex, formed from self-assembling subunits. The protein cage can reversibly form iron into a caged biomineral, Fe2C>3.H2O, in plants, animals, and bacteria. Iron oxy-biominerals inside the protein nanocages are iron concentrates for protein synthesis, and Fe(ll) / oxygen / peroxide traps (Fenton chemistry reactants) for antioxidant protection. The iron contained in ferritin is concentrated 100 billion times above the solubility of ferric ions in a nontoxic, accessible form. Ferritins occur in animals as approximately25 distinct isoforms depending on their proportions of the two primary subtypes of ferritins, H or L. These distinct subtypes differ in their tissue distribution, rates and mechanisms of iron oxidation, core formation and physiological iron turnover. Ferritin has many advantageous properties. For example, it can survive digestion largely intact, and is therefore more efficiently absorbed by the intestine than any other dietary iron source or iron supplement, because of the large amount of iron per ferritin molecule. Ferritin also survives treatment with high heat. The ferritin protein makes ferritin iron a naturally enteric coated, slow release, efficiently absorbed iron source. As such, ferritin may be used to supplement iron in humans or in animals in need of increased iron in their diet.

[0016] The present disclosure relates to the use of ferritin, specifically for its use in a cell culture medium, in lieu of transferrin. Transferrin is commonly used in cell culture media, such as KnockOut™ Serum Replacement (KSR). Use of ferritin in cell culture7105616786.2Attorney Docket No.: 116673-856389 media is new and unexpected, well beyond the existing knowledge that ferritin derived from plants and animals can be used as a dietary source for humans and other animals. Due to ferritin’s inherent stability and its low price (mainly related to plant- derived ferritin), the new use would revolutionize the cell culture industry, making cell culture media cost effective and practically feasible for mass production of food, feed or medical products, such as cultivated meat, biomass compositions or vaccine production. In the present disclosure, the ferritin used can be from any resource, such as from a plant, a bacterium, an alga, a fungus, or an animal. In one aspect, the ferritin comprises a recombinant ferritin. In another aspect, the ferritin used is from a plant, optionally a legume plant, such as soybean (Glycine max) or pea (Pisum sativum). In another aspect, the ferritin used in the cell culture media may have 24 subunits of L- and H-chains in varied proportions or may be any portion or segment thereof. In another aspect, the ferritin used in the cell culture is composed mostly, but not exclusively, of L-ferritin. In yet another aspect, the ferritin used in the cell culture is composed mostly, but not exclusively, of H-ferritin. In another aspect, the ferritin used in the cell culture is composed of a mixture of H- and L-ferritin in any proportion. In one aspect, the amount of ferritin used in the cell culture media is sufficient to support cell growth and cell survival. In another aspect, the ferritin in the cell culture medium is in an amount from about 50 pg / ml to about 150 pg / ml, optionally about 80 pg / ml to about 120 pg / ml, or preferably about 100 pg / ml.II. Cell Culture Medium

[0017] The present disclosure also relates to a cell culture medium. Cell culture media, also known as growth media, refers to an ex vivo nutritional environment, wherein cells grow, proliferate, and survive as they would in an in vivo biological system. Cell culture medium may be in a gel, liquid, or solid form. To support cellular growth ex vivo, a cell culture medium requires the same fundamental inputs as required in vivo: a mixture of a carbon-based energy source (e.g., glucose), amino acids, salts, vitamins, buffers, and other components to support cell viability and proliferation. An exemplary cell culture medium may comprise albumins, one or more amino acids, one or more vitamins, one or more transferrin, one or more antioxidants, one or more insulins or insulin substitutes, one or more collagen precursors, and one8105616786.2Attorney Docket No.: 116673-856389 or more trace elements. The composition of a cell culture medium is critical to ensure cell proliferation and survival. Compositions of cell culture media have been adjusted and optimized over time, and these adjustments and / or variations are often cell-type specific. For example, the composition of cell culture media suitable to grow stem cells is often different from formulations of media that support the growth of fibroblast or cancer cell lines. Similarly, growth media formulations are often specific for the species from which the cells were derived, where growth media used to propagate mammalian cells differs from growth media used to support avian cell lines. In one aspect, the cell culture medium disclosed herein can support the growth of mammalian stem cell growth. In another aspect, the cell culture medium disclosed herein can support growth of immortalized cell lines. In yet another aspect, the cell culture medium disclosed herein can support the growth of both mammalian and nonmammalian animal cell lines.Ila Serum

[0018] Serum used herein refers to an animal’s blood serum, which is a proteinrich liquid separated out from blood coagulates. In other words, serum is an animal ingredient. In one aspect, serum is derived from animal blood by removing blood cells and clotting proteins. Therefore, serum contains blood proteins except clotting factors, electrolytes, antibodies, antigens, hormones, and any exogenous substances. Serum does not contain all the formed elements of blood, which include blood cells, white blood cells (leukocytes, lymphocytes), red blood cells (erythrocytes), and platelets. It is noted that serum can support cellular growth and proliferation. Fetal bovine serum (FBS) is the commonly used animal serum supplement in a cell culture medium. The exact composition of FBS is variable depending at least upon its source, its extraction and preparation methods. It is generally accepted that FBS is a mixture of hormones, key proteins, lipids, growth factors, and trace elements that are essential for cell proliferation and / or survival. For example, FBS sourced from bovine fetuses contains more than 1 ,000 components such as growth factors, hormones, and transport proteins (including transferrin) that contribute to cell growth when supplemented into culture media.9105616786.2Attorney Docket No.: 116673-856389

[0019] FBS has many disadvantages, such as high cost and known issues for quality and reproducibility, animal welfare concerns, fraudulent marketing, and the lack of legal regulation in obtaining FBS from living calf fetuses. To overcome these advantages, various serum-free culture media have been developed. One example of serum-free cell culture medium is KnockOut™ Serum Replacement (KSR) culture medium, which comprises animal-ingredient, such as animal-derived albumin and / or transferrin. KSR can replace FBS in various cell cultures. KSR contains transferrin. In various aspects, the KSR used has the features disclosed in the manufacture’s website https: / / .thefmofisher.com / order / cataioq / product / 10828028, the contents of which are expressly incorporated by reference in its entity.

[0020] In one aspect, the present invention provides a serum-free, KSR-free cell culture medium. In another aspect, the present invention provides a serum-free, KSR- containing cell culture medium. In yet another aspect, the present invention provides a serum-free, animal-ingredient containing cell culture medium. In yet another aspect, the present invention provides a serum-free, animal-ingredient free cell culture medium. In yet another aspect, the present invention provides a serum-containing cell culture medium. The KSR-free or the KSR-containing cell culture medium comprises or is obtained by combining one or more ingredients selected from the group consisting of albumins or albumin substitutes, one or more amino acids, one or more vitamins, one or more transferrin or transferring substitutes, one or more antioxidants, one or more insulins or insulin substitutes, one or more inorganic salts, and one or more trace elements. lib Transferrin

[0021] Culture media, either serum-containing or serum replacement medium such as KSR, typically contain transferrin. Transferrin, like albumin, is commonly used in cell culture media. Transferrin is a group of proteins that assist in the control, transport, and delivery of ferric iron (Fe3+) to cells. Iron is involved in many cellular processes, serving as a cofactor for proteins and enzymes involved in metabolism, DNA synthesis and repair, and heme-mediated oxygen transport in vivo. Its levels within a cell must be tightly regulated, as deficiencies can lead to apoptosis and a surplus can lead to10105616786.2Attorney Docket No.: 116673-856389 toxic build-up of reactive oxygen species produced via the Fenton reaction. Transferrin mediates this regulation with two ultra-high affinity ferric ion binding pockets. As consecutive iron ions are bound, the affinity for the transferrin receptor increases. This permits two iron-bound transferrin to form a complex with their receptor, resulting in the delivery of the iron content to intracellular carriers such as ferritin. Iron-free transferrin (i.e., apotransferrin) can then be recycled to resume scavenging free iron in the serum or cell culture medium. Transferrin is highly abundant in Fetal Bovine Serum (FBS). FBS is a rich source of iron and protein- complexed iron, which increases in concentration throughout gestation. In the absence of serum or KSR, ferric and / or ferrous iron is typically delivered as a sulfate or nitrate salt in the basal media, and recombinant transferrin is added to regulate iron homeostasis and scavenge free iron to prevent oxidative damage.

[0022] The present invention provides a cell culture medium comprising ferritin in lieu of transferrin. In one aspect, the cell culture medium is called “ferritin-based cell culture medium”, in which the medium is substantially free of transferrin, indicating there may be a minimal amount or residual amount of transferrin in the medium, but the amount is not sufficient or adequate to support cell growth in the absence of additional exogenous transferrin. In one aspect, the ferritin-based cell culture medium comprises no animal ingredients. In another aspect, the ferritin-based cell culture medium comprises minimal amount of animal ingredients. In another aspect, the ferritin-based cell culture medium is animal-ingredient containing. An animal ingredient refers to any organ, tissue, cell, molecule, fluid, substance, chemical or other biological material isolated directly from a eukaryote. In one aspect, the ferritinbased cell culture medium is KSR-free and animal-ingredient free. In another aspect, the ferritin-based cell culture medium is KSR-containing but without the transferrin therein. KSR contains bovine-derived albumin and porcine-derived transferrin. In other words, KSR contains animal ingredients and is an animal-based product.

[0023] The ferritin-based cell culture medium may be substantially free of animal ingredients. In one aspect, the ferritin-based cell culture medium is plant-based, indicating all components in the culture medium are obtained or isolated from a plant, not from an animal. In another instance, the ferritin- based cell culture medium may be11105616786.2Attorney Docket No.: 116673-856389 selected from an animal ingredient free medium, serum-free medium, xeno-free medium, animal origin-free medium, and a chemically-defined medium. In such media, essential media components such as albumin may be replaced with recombinant plant-based components. Non-limiting examples of animal-free media include Essential 8™ medium, Beefy-9, B8, and ExCellerate™ iPSC Expansion Medium. Exemplary ingredients and respective concentrations in Essential 8™ and Beef-9 media are provided in Table 1 . In such instances, transferrin may be substituted with an equal, lower, or a different amount of ferritin. For example, the transferrin disclosed in Table 1 in the Essential 8™ medium and Beefy-9 media may be replaced with ferritin.Table 1 : Exemplary components of Essential 8™, Beefy-9, and B8 growth media12105616786.2Attorney Docket No.: 116673-856389

[0024] Any medium that is suitable for culturing cells for generating a biomass, for example cultivated meat, and with transferrin substituted with ferritin is contemplated for use as ferritin-based cell culture medium.

[0025] The amount of ferritin present in the ferritin-based cell culture medium may be at about 1 pg / ml to about 1500 pg / ml. In some instances, the ferritin may be present in the ferritin-based cell culture medium at about 1 pg / ml, about 5 pg / ml, about 10 pg / ml, about 20 pg / ml, about 30 pg / ml, about 40 pg / ml, about 50 pg / ml, about 60 pg / ml, about 70 pg / ml, about 80 pg / ml, about 90 pg / ml, about 100 pg / ml, about 150 pg / ml, about 200 pg / ml, about 250 pg / ml, about 300 pg / ml, about 350 pg / ml, about 400 pg / ml, about 450 pg / ml, about 500 pg / ml, about 550 pg / ml, about 600 pg / ml, about 700 pg / ml, about 750 pg / ml, about 800 pg / ml, about 850 pg / ml, about 900 pg / ml, about 950 pg / ml, about 1000 pg / ml, about 1150 pg / ml, about 1200 pg / ml, about 1250 pg / ml, about 1300 pg / ml, about 1350 pg / ml, about 1400 pg / ml, about 1450 pg / ml, about 1500 pg / ml, or more.He Inorganic Salts

[0026] A cell culture medium often contains inorganic salts. These inorganic salts may be composed of cations and anions that fully dissociate in solution. Some exemplary essential salts include calcium chloride, potassium chloride, magnesium sulfate, sodium chloride, sodium phosphate, sodium bicarbonate, as well as salts containing zinc, copper, and / or iron ions. In one aspect, the cell culture medium may contain NO iron supplement, and the iron presented may be in the form of association with ferritin. In another aspect, the cell culture medium may contain an iron supplement, in addition to the iron associated with ferritin. In some aspects, the iron supplement comprises iron (III) chloride, iron (II) chloride, iron (ll)-bisglycinate, or any combination thereof. Iron (ll)-bisglycinate contained in the cell culture medium may be in any amount or concentration sufficient to provide the support for cell growth105616786.2Attorney Docket No.: 116673-856389 and / or cell survival. In some aspect, the concentration may be from about 10 pg / ml to about 100 pg / ml. In one aspect, the iron supplement is about 37.26 pg / ml iron (II)- bisglycinate.

[0027] In a specific instance, the cell culture medium may not comprise an iron supplement. More specifically, the cell culture medium may not comprise NO iron supplement.

[0028] The present disclosure also encompasses a method of producing a ferritinbased cell culture medium. In one aspect, the method comprises: (i) obtaining or having obtained a commercially available cell culture medium; (ii) obtaining or having obtained ferritin, such as in a particle form; (iii) preparing the cell culture medium based on the manufacturer’s manual, except replacing transferrin with the ferritin, thereby obtaining a ferritin-based cell culture medium; and (iv) storing such ferritin-based cell culture medium or using it in cell culture. In one aspect, the prepared ferritin-based culture medium is substantially free of transferrin. In another aspect, the ferritin in particle form is obtained from a commercially available source. In one aspect, the ferritin is extracted from a plant or an animal tissue. In yet another aspect, the ferritin is obtained from a recombinant ferritin purchased from a commercially available source.

[0029] The present disclosure encompasses another method of producing a ferritin-based cell culture medium. In one aspect, the method comprises: (i) obtaining or having obtained ingredients comprising albumins or albumin substitutes, one or more amino acids, one or more vitamins, one or more antioxidants, one or more insulins or insulin substitutes, one or more inorganic salts, and one or more trace elements; (ii) obtaining or having obtained a ferritin, such as in a particle form; (iii) preparing the ferritin-based cell culture medium by combining the ingredients and ferritin in a gel or liquid carrier; and (iv) storing such ferritin-based cell culture medium or using it in cell culture. In one aspect, the prepared ferritin-based culture medium is substantially free of transferrin. In one aspect, the ferritin is obtained from a commercially available source. In one aspect, the ferritin is extracted from a plant or14105616786.2Attorney Docket No.: 116673-856389 animal tissue. In yet another aspect, the ferritin is obtained from a recombinant ferritin produced in house and / or purchased from a commercially available source.

[0030] Any cell suitable may be used in conjunction with the ferritin-based cell culture medium. Non-limiting examples of cells that may be cultured in the ferritinbased cell culture medium include stem cells, pluripotent stem cells, induced pluripotent stem cells (iPSC), embryonic stem cells (ESCs), immortalized cell lines, and primary cells. In an exemplary instance, the cells may be ESCs, for example mESC or iPSC. The cells may be human or non-human animal cells. Non-human animal cells include cells from cows, sheep, pigs, deer, roe deer, moose, kangaroo, duck, chicken, goose, fish, insects, and rodents such as mouse, rat, guinea pig or squirrel. Examples of cell line in use with the method include but not limited to DLEC, Mackl , CSG, ICF, Porcine_primary_CB, WAG, SAF-1 , ZEM2S, SICE, SIGE, Rabbit_tissue_BiolVT, Bovine_primary_Astarte, IEE, CIDuF, OnIL, Bovine_primary_CB, CCF, CMgM-1 , PHF, ZF4, BF-2, QM7, ZFiM-1 , PSF, IEG, Bovine_tissue_BiolVT, TTCF, SICH, Porcine_primary_Accegen, CSC-I9234L, Bovine_primary_Accegen, CHSE / F (formerly known as CHSE-214), FtGF, Porcine_tissue_BiolVT, and C2C12.

[0031] The cells may be cultured in the ferritin-based cell culture medium using any suitable method. For example, cells may be seeded in the ferritin-based cell culture. The conditions for cell culture may be optimized for maximal growth, proliferation, and / or differentiation of the cells. Alternatively, the cells may be cultured in a scaffold supplemented with ferritin-based cell culture, or may be transferred to a scaffold after growth, proliferation, and / or differentiation in the ferritin-based cell culture.III. Biomass Composition and Cultivated meat

[0032] The present disclosure encompasses a biomass composition. Biomass, as used herein refers collectively to the biological materials generated in a bioreactor, including but not limited to, cell culture medium, spent cell culture medium, cell lines, cells proliferated in the bioreactor and any metabolites, byproducts or debris produced during cell growth. Biomass compositions have many utilities. For example, biomass15105616786.2Attorney Docket No.: 116673-856389 from bacteria, yeasts, filamentous fungi, or microalgae - commonly referred to as microbial protein - is an alternative protein source that has long been proposed as a solution to food scarcity as it can be more sustainable than conventional food protein sources.

[0033] The present disclosure encompasses a cell biomass composition, wherein the composition is produced ex vivo by culturing a cell line in a culture medium comprising ferritin and substantially free of transferrin, i.e., the ferritin-based cell culture medium. In some aspects, a cell line may be cultured in the ferritin-based cell culture medium and may comprise a stem cell line, a fibroblast cell line, a cancer cell line, an immortalized cell line, or any combination thereof. The stem cell comprises an embryonic stem cell, an induced pluripotent stem cell, or any cell line combination thereof. The cancer cell line comprises a Hela cell line, a CHO cell line, or any cell line combination thereof. The immortalized cell line comprises a myoblast cell line, an adipocyte cell line, or any combination thereof. Other examples of cell lines include but not limited to DLEC, Mackl , CSG, IGF, Porcine_primary_CB, WAG, SAF-1 , ZEM2S, SICE, SIGE, Rabbit_tissue_BiolVT, Bovine_primary_Astarte, IEE, CIDuF, OnIL, Bovine_primary_CB, CCF, CMgM-1 , PHF, ZF4, BF-2, QM7, ZFiM-1 , PSF, IEG, Bovine_tissue_BiolVT, TTCF, SICH, Porcine_primary_Accegen, CSC-I9234L, Bovine_primary_Accegen, CHSE / F (formerly known as CHSE-214), FtGF, Porcine_tissue_BiolVT, C2C12, and any combination thereof. The cells may be human or non-human animal cells. Non-human animal cells include cells from cows, sheep, pigs, deer, roe deer, moose, kangaroo, duck, chicken, goose, fish, insects, and rodents such as mouse, rat, guinea pig or squirrel.

[0034] The biomass composition may be produced in a bioreactor. The bioreactor may have a volume in the range of about 500 mL to about 30,000 L, or more specifically from about 2 L to about 5,000 L. In an exemplary instance, the ferritinbased cell culture medium may be loaded into a bioreactor, and seeded with one or more cells. The bioreactor may comprise an agitator and one or more air spargers. The bioreactor may further comprise a structure or a scaffold for the growing the cells. The bioreactor may be maintained at a suitable temperature for growth, proliferation, and / or differentiation of the cells. For examples, the bioreactor may be maintained at16105616786.2Attorney Docket No.: 116673-856389 temperature of about 25° C to about 37° C. The cells may be incubated in the bioreactor for about 1 to about 30 days or more. The biomass composition is then generated by harvesting the cells, media, and media components. In one instance, the biomass composition comprises substantially everything harvested from the bioreactor. In a specific aspect, the biomass composition does not comprise cells separated from the medium by, for example, centrifugation.

[0035] In one aspect, the biomass composition contains ferritin and is substantially free of transferrin. In some aspects, the amount of ferritin used is sufficient to support cell growth and cell survival. In one aspect, the amount of ferritin used is highly dependent on the cell line being cultured. For example, the amount of ferritin in the composition is different when growing stem cells than when growing fibroblast cells. In another aspect, the ferritin in the biomass composition is in an amount from about 50 pg / ml to about 150 pg / ml, optionally about 80 pg / ml to about 120 pg / ml, or preferably about 100 pg / ml.

[0036] The present disclosure also encompasses a biomass composition further comprising a functional ingredient to supplement the composition, including but not limited to supplement composition’s protein content, lipid content, sugar content, fiber content, calcium content, iron content, or any combination thereof. In one aspect, the functional ingredient comprises, including but not limited to, a flavorant, a colorant, a preservative, an antioxidant. In one aspect, the functional ingredient comprises food grade, feed grade, lab grade, pharmaceutical grade, or chemical grade. In one aspect, the composition further comprises a food grade or feed grade flavorant, a food grade or feed grade colorant, a food grade or feed grade preservative, a food grade or feed grade antioxidant, or any combination thereof. In another aspect, the cell biomass composition further comprises food grade or feed grade flavorant in a chicken flavorant, a beef flavorant, a pork flavorant, a mouse flavorant, a fish flavorant, a smoke meat flavorant, or any combination thereof.

[0037] In another aspect, the biomass composition is incorporated or integrated into a food product, a feed product, a veterinary product, a dietary supplement product, a performance-enhancing product, a nutraceutical product, a chemical product, or a17105616786.2Attorney Docket No.: 116673-856389 pharmaceutical product. In yet another aspect, the biomass composition is used as a culture medium component, as a carrier component, as an excipient component, as a reagent, or any combination thereof.

[0038] In yet another aspect, the ferritin-based cell culture medium is used as a culture medium for culturing and / or proliferating cells to generate a cultivated meat product. Cells, for example, stem cells, may be cultured in the ferritin-based cell culture medium. Following proliferation, the stem cells spontaneously differentiate. The cells then may be separated for use as cultivated meat. In a specific aspect, the cells are not separated from the culture medium for use as cultivated meat. In such instances, the cells, and the soluble animal-derived nutrients synthesized by the cells are extruded into the surrounding medium. The cells and the medium are then harvested and used as an ingredient in food, for example, pet food. In a specific aspect, cultured cells and surrounding nutrient-rich culture, i.e., the biomass composition comprising protein- and fat-containing cells that were proliferated from pluripotent stem cells from a genetically-stable cell line, for example, a mouse cell line, derived under sterile conditions are used. Cell line may be confirmed to be free of pathogens and contaminating material. The culture medium at the time of harvest may comprise a) exogenous non-animal derived food- or feed-grade ingredients that were fed to the cells; and / or b) endogenous animal-based nutrients produced by the same cells and secreted into the surrounding liquid environment.

[0039] Any cell line suitable for production of cultivated meat, generally known in the art may be used. For example, embryonic stem cell, an induced pluripotent stem cell, or any cell line combination thereof, may be used to produce the cultivated meat. Other examples of cell lines include but not limited to DLEC, Mackl , CSG, ICF, Porcine_primary_CB, WAG, SAF-1 , ZEM2S, SICE, SIGE, Rabbit_tissue_BiolVT, Bovine_primary_Astarte, IEE, CIDuF, OnIL, Bovine_primary_CB, CCF, CMgM-1 , PHF, ZF4, BF-2, QM7, ZFiM-1 , PSF, IEG, Bovine_tissue_BiolVT, TTCF, SICH, Porcine_primary_Accegen, CSC-I9234L, Bovine_primary_Accegen, CHSE / F (formerly known as CHSE-214), FtGF, Porcine_tissue_BiolVT, C2C12, and any combination thereof. The cells may be human or non-human animal cells. Non-human animal cells include cells from cows, sheep, pigs, deer, roe deer, moose, kangaroo,18105616786.2Attorney Docket No.: 116673-856389 duck, chicken, goose, fish, insects, and rodents such as mouse, rat, guinea pig or squirrel.

[0040] The present disclosure further encompasses a cultivated meat product containing the cells cultured from the ferritin-based culture media. Cultivated meat is a form of cellular agriculture where meat is produced by culturing animal cells in vitro or ex vivo. Cultivated meat may also be called cultured meat, healthy meat, slaughter- free meat, vat-grown meat, lab-grown meat, cell-based meat, and synthetic meat, among others. Cultured meat is often produced using tissue engineering or bioengineering techniques. Since it relies on cell culture, the composition, quality, and productivity of cell culture medium plays a critical role in cultivated meat production. Traditional transferrin-containing culture medium has limited practical use, due to its high cost.

[0041] The present disclosure provides a cultivated meat composition wherein the cells are grown from a ferritin-based culture medium. Ferritin costs significantly less than transferrin (roughly US$0.022 / L vs roughly US$60 / L, respectively, in current market), therefore ferritin-based culture medium is much more affordable than its transferrin counterparts. Consequently, ferritin-based culture medium would make growing cells in bioreactors cost effective and practically feasible for mass production, one giant step closer to leveraging cultivated meat to solve world famine.

[0042] In one aspect, the cell lines used to produce cultivated meat include among others, stem cells, fibroblast cells and / or immortalized cell lines. The stem cells comprise at least embryonic stem cells and induced pluripotent stem cells, or any combination thereof. The immortalized cell lines comprise myoblast cell lines, adipocyte cell lines, or any combination thereof. In one aspect, the cultivated meat composition further comprises a functional ingredient to supplement the product’s protein content, lipid content, sugar content, fiber content, calcium content, iron content, or any combination thereof. In another aspect, the cultivated meat composition further comprises a food grade flavorant, a food grade colorant, a food grade preservative, a food antioxidant, or any combination thereof. In yet another aspect, the cell biomass composition contains a food grade flavorant comprising a19105616786.2Attorney Docket No.: 116673-856389 chicken flavorant, a beef flavorant, a pork flavorant, a fish flavorant, a smoke meat flavorant, or any flavorant combination thereof.

[0043] The present disclosure encompasses a method of producing a cultivated meat composition. The method comprises as shown partially in FIG.4A: (i) obtaining or having obtained a cell line suitable for cultivated meat production; (ii) culturing the cell line in a transferrin-based culture medium; (iii) weaning cultured cells off the transferrin-based culture medium and into a fresh ferritin-based culture medium; (iv) passaging the cultured cells into the fresh ferritin-based culture medium for one or more passage; and (v) collecting the cultured cells, the cultured cells together with the spent media, or the full biomass composition, thereby producing the cultivated meat composition. In one aspect, the transferrin-based culture medium is substantially free of ferritin; and the ferritin-based culture medium is substantially free of transferrin. In one aspect, the weaning step comprises transferring and culturing the cells in a mixed culture medium. The mixed culture medium comprises a mixture of the transferrin- based culture medium and a ferritin-based culture medium, wherein the transferrin- based culture medium accounts for about 30% (v / v) to about 50% (v / v) of the mixed culture medium. In one aspect, the mixture contains about 50% transferrin-based culture medium and 50% ferritin-based culture medium. In another aspect, weaning steps may be repeated 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10 or more times. In another aspect, the passaging step is repeated 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10 or more times. Alternatively, as shown in FIG. 4B, in step (iii) cultured cells are weaned into a mixture of transferrin- based medium, which comprises about 50% of KSR standard transferrin-containing medium, and about 50% recombinant transferrin-based medium. Then followed by another passage to 100% recombinant transferrin-based medium. In one aspect, the ferritin-based culture medium may be the same or different in the weaning or the passaging steps. For example, in weaning round 1 , the ferritin-based medium contains plant-derived ferritin; in passaging round 1 , the ferritin-based medium contains recombinant ferritin. Or in weaning round 1 , the ferritin-based medium contains animal derived or recombinant ferritin; in passaging round 2 and thereafter, the ferritin-based medium contains plant-derived ferritin. FIGs. 4A-4B merely provide exemplary uses of the ferritin-based media and should not be interpreted as20105616786.2Attorney Docket No.: 116673-856389 exhaustive. In another aspect, the transferrin-based culture medium is the KSR medium and is prepared based on the manufacture’s user manual. In another aspect, the transferrin-based culture medium is prepared in house which is KSR-free and animal-ingredient free. In one aspect, the ferritin-based medium comprises ferritin derived from an animal, derived from a plant, a recombinant ferritin, or any combination thereof. In one aspect, the ferritin-based medium comprises ferritin derived from a plant. In one aspect, such plant comprises Glycine max or Pisum sativum. In one aspect, the ferritin-based is prepared by using KSR medium as the base and replaces transferrin therein with ferritin. In one aspect, the replacement results in a culture medium substantially free of transferrin. In one aspect, the replacement results in a culture medium containing less than about 10% of transferrin (v / v). In one aspect, the replacement results in a culture medium containing less than about 8% of transferrin (v / v). In one aspect, the replacement results in a culture medium containing less than about 5% of transferrin (v / v). In one aspect, the replacement results in a culture medium containing less than about 3% of transferrin (v / v). In one aspect, the replacement results in a culture medium containing less than about 2% of transferrin (v / v). In one aspect, the replacement results in a culture medium containing less than about 1% of transferrin (v / v). In one aspect, the replacement results in a culture medium containing less than about 0.5% of transferrin (v / v). In one aspect, the replacement results in a culture medium containing less than about 0.2% of transferrin (v / v). In one aspect, the replacement results in a culture medium containing less than about 0.1 % of transferrin (v / v). In another aspect, the ferritin-based medium is prepared by incorporating various ingredients. Such ingredients include but are not limited to, albumins or albumin substitutes, one or more amino acids, one or more vitamins, one or more ferritins, one or more antioxidants, one or more insulins or insulin substitutes, one or more inorganic salts, and one or more trace elements. For example, the preparation method comprises: (i) obtaining or having obtained ingredients of albumins or albumin substitutes, one or more amino acids, one or more vitamins, one or more antioxidants, one or more insulins or insulin substitutes, one or more inorganic salts, and one or more trace elements.; (ii) obtaining or having obtained a ferritin in a particle form; (iii) preparing the ferritin-based cell21105616786.2Attorney Docket No.: 116673-856389 culture medium by combining the ingredients and ferritin in a gel or liquid carrier; and (iv) storing such ferritin-based cell culture medium or using it in cell culture.

[0044] The weaning or transfer of cells may be performed after culturing the cells in the transferrin-based culture medium or the mixed culture medium for about 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 or more days.

[0045] The mixed culture medium may comprise a transferrin-based culture medium and a ferritin-based culture medium. In some instances, the transferrin-based culture medium in the mixed medium may be at about 1 %, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more. In further instance, the ferritin-based culture medium in the mixed medium may be at about 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more. In a specific instance, the first step of weaning or transfer of cells may comprise a 50% transferrin-based culture medium and 50% ferritin-based culture medium to generate a first mixed culture medium. The second step of weaning or transfer of cells may comprise replacing 50% of the first mixed culture medium with ferritin based culture medium to generate a second mixed culture medium. 50% of the second mixed culture medium may be replaced by ferritin-based culture medium in subsequent step. This may be continued until the cell culture medium is full weaned of transferrin, i.e., comprises 0% transferrin based medium and 100% ferritin based culture medium.

[0046] In one instance, the transferrin content in a transferrin based medium may be about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11 %, about 12%, about 13%, about 14%, about 15% or more. In a specific aspect, the transferrin content may be about 10%.

[0047] The transferrin content in a first mixed medium, i.e., 50% transferrin-based culture medium and 50% ferritin-based culture medium may be about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10%. In a specific aspect, the first mixed medium may comprise about 5% transferrin.22105616786.2Attorney Docket No.: 116673-856389

[0048] The subsequent mixed medium i.e., weaning by replacement with ferritin based medium may comprise about less than 1%, about 1 %, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10% transferrin. Such medium may comprise, in a specific instance, less than 1% transferrin.

[0049] A transferrin devoid medium, obtained after the complete weaning process may be substantially devoid of transferrin. In such instances, the medium may comprise less than about 0.1%, about 1%, about 2%, about 3%, about 4%, about 5%, or lesser amount of transferrin. In some instances, the medium may comprise carry over transferrin. Such carry over transferrin may be about less than 0.1%, about 1 %, or about 2%.

[0050] The present disclosure encompasses another method of producing a cultivated meat composition. The method comprises: (i) obtaining or having obtained a cell line suitable for cultivated meat production; (ii) culturing the cell line in a ferritinbased culture medium; (iii) passaging the cultured cells into another and / or same fresh ferritin-based culture medium for one or more passage; and (iv) collecting the cultured cells or the cultured cells together with the spent media, thereby producing the cultivated meat composition; wherein the ferritin-based culture medium is substantially free of transferrin. In one aspect, the ferritin-based medium comprises ferritin derived from an animal, derived from a plant, a recombinant ferritin, or any combination thereof. In one aspect, the ferritin-based medium comprises ferritin derived from a plant. In one aspect, such plant comprises Glycine max or Pisum sativum. In one aspect, the ferritin-based is prepared by using KSR medium as the base and replaces transferrin therein with ferritin. In one aspect, the replacement results in a culture medium substantially free of transferrin. In one aspect, the replacement results in a culture medium containing less than about 10% of transferrin (v / v). In one aspect, the replacement results in a culture medium containing less than about 8% of transferrin (v / v). In one aspect, the replacement results in a culture medium containing less than about 5% of transferrin (v / v). In one aspect, the replacement results in a culture medium containing less than about 3% of transferrin (v / v). In one aspect, the replacement results in a culture medium containing less than about 2% of transferrin23105616786.2Attorney Docket No.: 116673-856389(v / v). In one aspect, the replacement results in a culture medium containing less than about 1 % of transferrin (v / v). In one aspect, the replacement results in a culture medium containing less than about 0.5% of transferrin (v / v). In one aspect, the replacement results in a culture medium containing less than about 0.2% of transferrin (v / v). In one aspect, the replacement results in a culture medium containing less than about 0.1% of transferrin (v / v). In another aspect, the ferritin-based is prepared by incorporating various ingredients. Such ingredients include but are not limited to, albumins or albumin substitutes, one or more amino acids, one or more vitamins, one or more ferritins, one or more antioxidants, one or more insulins or insulin substitutes, one or more inorganic salts, and one or more trace elements. For example, the preparation method comprises: (i) obtaining or having obtained ingredients of albumins or albumin substitutes, one or more amino acids, one or more vitamins, one or more antioxidants, one or more insulins or insulin substitutes, one or more inorganic salts, and one or more trace elements.; (ii) obtaining or having obtained a ferritin in a particle form; (iii) preparing the ferritin-based cell culture medium by combining the ingredients and ferritin in a gel or liquid carrier; and (iv) storing such ferritin-based cell culture medium or using it in cell culture.

[0051] The biomass composition and / or the cultivated meat generated by the methods disclosed herein may be incorporated into a food or a feed product. In one aspect, the biomass composition and / or the cultivated meat may be processed before incorporating into the food or feed product. For example, the biomass composition and / or the cultivated meat may be processed by at least one product processing device selected from a mixer, a grinder, a chopper, a lyophilizer, a steamer, a blender, a cooker, a boiler, a dryer, a vacuum dryer, a grill, a roaster, a washing device, a reaction vessel, a bioreactor; a filtration device, a centrifuge, a sieve, a grill, a heater, UV lamp, IR lamp, extruder, chiller, freezer; and / or any other product processing device. The biomass composition and / or the cultivated meat may be mechanically and / or chemically homogenized to disrupt any clumps, aggregates, and lumps that may form during cell culture. The biomass composition and / or the cultivated meat may be inactivated to stop proliferation, differentiation, maturation, any cell metabolic processes. Non-limiting examples of inactivation include drying, chemical detergent24105616786.2Attorney Docket No.: 116673-856389 induced lysis, cooling and / or osmotic shock. Alternatively, the biomass composition and / or the cultivated meat may not be further processed before incorporating into the food or feed product.Illa Food or Feed Products

[0052] A product is an item ready to be consumed or used without further (or requiring minimal) processing or modification. A food product refers to a product suitable for consumption by a human being, whereas a feed product refers to a product suitable for consumption by an animal. Because they are intended for human or animal consumption, all components or ingredients in the products would be at least food or feed grade. Food grade means that a material is fit for human consumption or permitted to come in contact with food. For example, in the U.S., a material considered GRAS (Generally Recognized as Safe) may be used in a food product. In another aspect, food grade also limits on microbial count such as in a GMP environment. For example, food ingredients often have to have less than 0.5% impurity. Both food and feed products are aimed to provide humans and / or animals with the required nutrition, rather than for therapeutic or treatment use. For a food product, the human being includes an infant, a toddler, a youth, a middle-aged, or an elderly. Such food products also include food supplements, dietary supplements, or special foods, such as baby formula, diabetic food, weight-loss food. For a feed product, also called a “veterinary product”, the animal can be a mammal or a non-mammal. The animal can be a pet or a livestock. Pets include, but not limited to, dogs, cats, fish, rabbits, pigs, guinea pigs, ferrets, birds, or reptiles. Livestock includes, but not limited to, chickens, pigs, cows, fish, and other farm animals. In one aspect, the feed product is integrated into a product that is nutritionally complete and balanced. In another aspect, the feed product is supplemented with other feed products to provide the animal with a complete diet so that they can live a healthy life. In another aspect, the feed product is fed to the animal as a dietary supplement or treats. Yet in another aspect, the present disclosure expressly encompasses animal treats that are often highly flavored or have a special chewy texture. In yet another aspect, the feed product is aimed at providing an enjoyable experience for the animals and may not be necessarily well balanced nutritionally. Further, in the present disclosure, a food or feed product also25105616786.2Attorney Docket No.: 116673-856389 expressly encompasses products commonly termed as dietary supplement products, specialty food, or performance-enhancing products. These products are designed to be consumed by either human beings or by animals.

[0053] In one aspect, the food or pet product of the present disclosure is free of animal ingredients. Yet in another aspect, the product contains animal ingredients. An animal ingredient refers to any organ, tissue, cell, molecule, or biological material isolated directly from an animal, but does NOT include the recombinant counterparts produced through bioengineering or other ex vivo methods. In one aspect, the composition is serum-free and KSR-free. In another aspect, the product is serum-free and KSR-containing. Serum refers to an animal’s blood serum, which is a protein-rich liquid separated out from blood coagulates. In other words, serum is an animal ingredient. In one aspect, the product is plant-based, indicating all components in the biomass are obtained or isolated from a plant, not an animal. In one aspect, such food or pet product is for oral consumption. In another aspect, such food or pet product is in the form of powder, capsules or tablets. In yet another aspect, the food or pet product could NOT be labelled as protein rich or protein enhanced or boosted.

[0054] A food product or a pet food may be produced by combining the cultivated meat or biomass composition with another food grade ingredient. The food grade ingredient that can be used in combination with the cultivated meat or biomass composition can include glucose and fructose, chicory root extract, inulin, starch, maltodextrin, lactose, maltose, sucrose and saccharose, rice, corn, potatoes, sweet potatoes, barley, oats, peas, soy, tapioca, lentils, chickpeas, sorghum, quinoa, millet, wheat, cassava, yams, pumpkin, carrots, beet pulps, apples, bananas, blueberries, cranberries, apricots, butternut squash, carrageenan, spirulina, pectin, pineapple, tomatoes, elderberries, rosehips, beets, celery, resistant starch, inulin, xanthan gum, cereals, grains, beta-glucans, psyllium, oat bran, what bran, cellulose, broccoli, cauliflower, guar gums, chicory roots, cranberries, squash, beans, group of waxy rice starch, waxy barley starch, waxy maize starch, waxy wheat starch, waxy potato starch, oat starch, gluten, sorbitol, spinach, grape, glycerol, soybean hulls, whole grain oat, grape, celery, olive oil, coconut oil, avocado oil, canola oil, sunflower oil, flaxseed oil, sesame oil, rapeseed oil, flaxseed oil, vegetable oils, corn oil, soy oil, cottonseed oil,26105616786.2Attorney Docket No.: 116673-856389 palm oil, linseed oil, menhaden oil, peanut oil, olestra, almonds, walnuts, cashews, pecans, macadamia nuts, hazelnuts, flaxseeds, sunflower seeds, pumpkin seeds, hemp seeds, sesame seeds, avocado, olives, almond butter, cashew butter, seaweed, tahini, hummus, lauric acid, linoleic acid, babassu oil, palmitoleic acid, cohune oil, palm kelner oil, tucum oil, soybean oil and any combination thereof. Other food ingredients used in combination with the cultivated meat or biomass composition may be vitamins, minerals, flavor imparting compounds, food color, digestibility enhancer, palatability enhancers, yeast extracts, bacterial extracts, and texture additives.

[0055] The pet food product may be a wet food product or a dry food product. The pet food product may be a snack, kibble, pate, chunks, pouch, gravy, a canned food. The pet food product may comprise about 0.1 % wt, about 1 wt% to about 99% wt% of the biomass composition or the cultivated meat. For example, the pet food may comprise about 1 wt%, about 5 wt%, about 10 wt%, about 20 wt%, about 30 wt%, about 40 wt%, about 50 wt%, about 60 wt%, about 70 wt%, about 80 wt%, about 90 wt%, or about 99 wt% biomass composition or the cultivated meat.

[0056] The biomass composition or the cultivated meat may be used as a replacement for one or more animal ingredient such as chicken, beef, salmon etc. as well as for chicken by-product, beef by-product and salmon by-product in a pet food product. In such aspects, the animal ingredient may be substituted with the biomass composition or the cultivated meat at a ratio of about 1 :1 , about 1 :2, about 1 :3, about 1 :4, about 1 :5, about 1 :6, about 1 :7, about 1 :8, about 1 :9, about 2:1 , about 3:1 , about 4:1 , about 5:1 , about 6:1 , about 7:1 , about 8:1 , about 9:1 , or about 10:1.

[0057] The food product also may be a product for human consumption. In such instances, the biomass composition or the cultivated meat may be used to produce or incorporated into food products non-limiting examples of which include bacon, ham, hotdog, prosciutto, sausage, burger, ground meat, patties, meatballs, jerky strips, nuggets, strips, meat loaf, meat cake forms, canned seasoned meat, sliced meat, salami, bologna, lunch meat, and the like. In one aspect, the meat products may be a plant-based meat alternative, or an animal-free product. The food product may comprise about 1 wt% to about 99% wt% of the biomass composition or the cultivated27105616786.2Attorney Docket No.: 116673-856389 meat. For example, the food product may comprise about 0.1 % wt%, about 1 wt%, about 5 wt%, about 10 wt%, about 20 wt%, about 30 wt%, about 40 wt%, about 50 wt%, about 60 wt%, about 70 wt%, about 80 wt%, about 90 wt%, or about 99 wt% biomass composition or the cultivated meat.Illb Pharmaceutical Product

[0058] The present disclosure encompasses a pharmaceutical product comprising the biomass composition. A pharmaceutical product is a product used to diagnose, cure, treat, or prevent a disease, or a product to relieve or alleviate symptoms from a disease. A pharmaceutical product often has two distinctive components: a pharmaceutical active ingredient (API) and an excipient suitable to carry the API to its site of action. An API is any ingredient that provides biologically active or other direct effect in the diagnosis, cure, mitigation, treatment, or prevention of disease or to affect the structure or any function of the body of humans or animals. In one aspect, the biomass encompassed by the present disclosure is the API for treating or preventing a disease. In one aspect, such disease may be associated with malnutrition or iron deficiency. Iron deficiency may result in anemia, i.e. , too few healthy red blood cells in the body. Malnutrition may manifest as physical or mental disability.I He Nutraceutical Product

[0059] A nutraceutical product refers to a food product or part of a food product that offers medical and / or health benefits in prevention or treatment of disease but could not be claimed as curative or therapeutic. Nutraceutical products often contain isolated and / or enhanced nutrients. They are often closely related to genetically engineered designer foods, functional foods, herbal products and processed foods such as cereal, soup and beverages. Functional food refers to foods that include any modified food or food ingredients that may provide a health benefit beyond the traditional nutrients. Nutraceutical products include food and / or feed for human or veterinary use, respectively. It may take many forms, such as food bars, chewing gums, drinks, fortified drinks, add-on powder supplements, tablets; lozenges, capsules, candies, and the like.28105616786.2Attorney Docket No.: 116673-856389IV. Effective Amount and / or Concentrations

[0060] The present disclosure provides ferritin in different amounts or concentrations, as the amount of ferritin required to support cell growth and cell survival is highly cell line dependent. For example, the amount of ferritin in the ferritinbased culture medium is different when growing stem cells than when growing fibroblast cells. Further, the amount of ferritin in the culture medium, in the biomass composition or in the cultivated meat composition would also be different and each can be measured by weight or weight concentration (w / w) or by volume (ml) or volume concentration (w / l). Generally speaking, the essential role of ferritin in these various compositions is to support cell growth and survival, therefore the required amount would follow a similar scheme. When measured in w / v, the ferritin required may be up to 200 pg / ml. In another aspect, the ferritin is in an amount from about 50 pg / ml to about 150 pg / ml, optionally from about 80 pg / ml to about 120 pg / ml, from about 90 pg / ml to about 110 pg / ml, or more preferably about 100 pg / ml. For example, the range of about 80 pg / ml to about 120 pg / ml encompasses about 80 pg / ml, about 82 pg / ml, about 84 pg / ml, about 85 pg / ml, about 86 pg / ml, about 88 pg / ml, about 89 pg / ml, about 90 pg / ml, about 92 pg / ml, about 94 pg / ml, about 95 pg / ml, about 96 pg / ml, about 98 pg / ml, about 100 pg / ml, about 102 pg / ml, about 104 pg / ml, about 105 pg / ml, about 106 pg / ml, about 108 pg / ml, about 110 pg / ml, about 112 pg / ml, about 114 pg / ml, about 115 pg / ml, about 116 pg / ml, about 118 pg / ml, about 120, and all the other concentration points in between. On the other hand, when measured in w / v, the ferritin required may be from about 10% to about 60% by weight. For example, ferritin may be present in an amount of from about 10% to about 15%, from about 15% to about 20%, from about 20% to about 25%, from about 25% to about 30%, from about 30% to about 35%, from about 35% to about 40%, from about 40% to about 45%, from about 45% to about 50%, or from about 55% to about 60% by weight of the total composition either in the culture medium, in the biomass or in the cultivated meat.Definitions29105616786.2Attorney Docket No.: 116673-856389

[0061] Unless defined otherwise, all technical and scientific terms used herein have the meaning commonly understood by a person skilled in the art to which present disclosure belongs. The following references provide one of skill with a general definition of many of the terms used in present disclosure: Singleton et al., Dictionary of Microbiology and Molecular Biology (2nd ed. 1994); The Cambridge Dictionary of Science and Technology (Walker ed., 1988); The Glossary of Genetics, 5th Ed., R. Rieger et al. (eds.), Springer Verlag (1991); and Hale & Marham, The Harper Collins Dictionary of Biology (1991). As used herein, the following terms have the meanings ascribed to them unless specified otherwise.

[0062] When introducing elements of the present disclosure or the preferred aspects(s) thereof, the articles “a”, “an”, “the” and “said” are intended to mean that there are one or more of the elements. The terms “comprising”, “including” and “having” are intended to be inclusive and mean that there may be additional elements other than the listed elements.

[0063] The term “comprising” means “including, but not necessarily limited to”; it specifically indicates open-ended inclusion or membership in a so-described combination, group, series and the like. The terms “comprising” and “including” as used herein are inclusive and / or open-ended and do not exclude additional, unrecited elements or method processes. The term “consisting essentially of’ is more limiting than “comprising” but not as restrictive as “consisting of.” Specifically, the term “consisting essentially of” limits membership to the specified materials or items and those that do not materially affect the essential characteristics of the present disclosure.

[0064] As used herein, the term “about” means 10% plus and / or minus of the recited amount or concentration. The amount includes weight or volume amount. The concentration includes weight (w / w) or volume concentrations (w / v). As used herein, term “substantial” means a material amount capable of causing the desired results. And term “substantially free” shall mean any amount less than a material amount that could not have caused the desired effect. Specifically for the amounts of ferritin and transferrin, the term “substantially free” would mean less than about 10% (w / w) or 10%30105616786.2Attorney Docket No.: 116673-856389(w / v) of the excluded form and preferably less than about 5% and more preferably less than about 2%, 1 %, and in some cases less than about 0.5% or 0.1 % w / w or w / v concentration.

[0065] As used herein, the terms “disease” or “disorder” are used interchangeably in the present disclosure. They refer to any condition, disorder or disease manifested as one or more physiological, physical and / or psychological symptoms or dysfunctions for which treatment is desirable, and includes previously and newly identified diseases or disorders on any organs, tissues or biological activities. As used herein, the term “pharmaceutical product” is a product designed to restore, remedy, or preserve health or wellbeing of a human being or an animal.

[0066] The terms “or” and “and / or,” as used herein, are to be interpreted as inclusive or meaning any one or any combination. Therefore, “A, B or C” or “A, B and / or C” mean any of the following: “A,” “B” or “C”; “A and B”; “A and C”; “B and C”; “A, B and C.” An exception to this definition will occur only when a combination of elements, functions, steps or acts are in some way inherently mutually exclusive.

[0067] The term “about”, as used herein, may refer to variation of a numerical quantity that can occur, for example, through typical measuring techniques and equipment, with respect to any quantifiable variable, including, but not limited to, mass, volume, time, distance, and amount. Further, given solid and liquid handling procedures used in the real world, there is certain inadvertent error and variation that is likely through differences in the manufacture, source, or purity of the ingredients used to make the compositions or carry out the methods and the like. The term “about” also encompasses these variations, which can be up to ± 5%, but can also be ± 4%, 3%, 2%,1%, etc.

[0068] For recitation of numeric ranges herein, each intervening number at the same decimal level there between (e.g., the same degree of precision) is explicitly contemplated. For example, for the range of 6-9, the numbers 6, 7, 8 and 9 are explicitly contemplated, and for the range 6.0-7.0, the numbers 6.0, 6.1 , 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6,9, and 7.0 are explicitly contemplated.31105616786.2Attorney Docket No.: 116673-856389

[0069] The term "pluripotent stem cells (PSCs)" refers to cells that can propagate indefinitely, as well as give rise to every other cell type in the body, including muscle cells, bone cells and fat cells.

[0070] The term "induced pluripotent stem cells (iPSCs)" refers to a type of pluripotent stem cell that can be generated directly from differentiated cells by reprogramming processes well known in the art.

[0071] The term "embryonic stem cells (ESC)" refers to a type of pluripotent stem cell derived from blastocyst.

[0072] The term “immortalized cell lines” refers to cells that, due to mutations, do not undergo replicative senescence and can be maintained in culture for long periods of time.

[0073] The term “primary cells” refers to cells freshly isolated from an animal tissue and grown in vitro.

[0074] The term “reprogramming” or “differentiating” refers to conversion of one specific cell type to another.

[0075] The publications discussed above are provided solely for their disclosure before the filing date of the present application. Nothing herein is to be construed as an admission that the present disclosure is not entitled to antedate such disclosure by virtue of prior disclosure.Examples

[0076] The following examples are included to demonstrate the disclosure. It should be appreciated by those of skill in the art that the techniques disclosed in the following examples represent techniques discovered by the inventors to function well in the practice of the disclosure. Those of skill in the art should, however, in light of the present disclosure, appreciate that many changes could be made in the disclosure and still obtain a like or similar result without departing from the spirit and scope of the32105616786.2Attorney Docket No.: 116673-856389 disclosure, therefore all matter set forth is to be interpreted as illustrative and not in a limiting sense.Example 1. Study Design

[0077] The present disclosure relates to various tests and experiments conducted on ferritin. Unless otherwise indicated, ferritin used in all studies was a food-grade ferritin isolated from plants. The ferritin contains >5% iron and is in particle form with 98% through USA #60 sieve. Tests were conducted both in suspension and adherent cultures. Exemplary instrumentations and tools were listed in Table 2.TABLE 2: Instrumentation and Tools Used in ExamplesTable 3: Preparation of Standard KSR Medium33105616786.2Atorney Docket No.: 116673-856389Example 2. Animal-Ingredient Free Ferritin-Based Medium for Culturing Stem Cells

[0078] A standard KnockOut™ Serum Replacement (KSR) culture medium was prepared according to Table 3. Additionally, a ferritin-based, animal-ingredient free, proprietary medium was prepared, and a transferrin-based, animal-ingredient free, proprietary medium was prepared. It was calculated that the animal-ingredient free ferritin-based medium contained about 100 pg / ml of ferritin and no detectable amount of transferrin. Cells from a mouse embryonic stem cell (mESC) line, preserved in cryovials, were retrieved and thawed in standard KSR medium in ABLE bioreactors and cultured at 37°C and 5% CO2. Passaging was performed using a combination of enzymatic and mechanical dissociation strategies. In order to wean to 100% ferritinbased medium, two different approaches were tested. In the first approach, after first passaging, cells were inoculated to a medium containing about 50% standard KSR medium and about 50% ferritin-based medium and cultured for 48 hours at 37°C and 5% CO2. After a second passaging, cells were inoculated in 100% ferritin-based medium. In the second approach, after the first passaging, cells were inoculated in a medium containing about 50% standard KSR medium and about 50% transferrin- based, animal-ingredient free, proprietary medium. After the second passage, cells34105616786.2Attorney Docket No.: 116673-856389 were inoculated to 100% transferrin-based, animal-ingredient free, proprietary media, and after the third passage, cells were inoculated to 100% ferritin-based, animalingredient free, proprietary media. In both approaches, mESCs were successfully proliferated in the serum-free and animal-ingredient free culture medium containing ferritin in lieu of transferrin. The mESCs obtained after passage 12 were characterized. Viability and number of cells were evaluated via Vial -Cassettes and NucleoCounter, respectively. As shown in FIG. 1 , ferritin versus transferrin provided an improved doubling time of mESCs in animal-ingredient free cell culture at passage 12. FIG. 2 showed ferritin yielded the same viability as transferrin for mESCs grown in animal-ingredient free cell culture at passage 12. Cell morphology was captured with the aid of a MikrOkular Teleskop Kamera through a microscope using 10x magnification, and presented in FIGs. 3A-3B, showing equivalent morphology of mESCs in cell culture with ferritin (FIG. 3A) versus cell culture with transferrin (FIG. 3B). These results demonstrated that ferritin can successfully replace transferrin in cell culture medium. FIGs. 4A-4B provided illustrative flow on how cells were weaned (transferred) and passaged to the ferritin-based medium.

[0079] Pluripotency of the cultured mESCs is evaluated using a quantitative PCR (qPCR). Further, differentiation is induced by removing Leukemia Inhibitory Factor (LIF) from the cultured mESCs and thus obtained mESCs are evaluated for pluripotency using qPCR.Example 3. Animal-Ingredient Free Ferritin-Based Medium for Culturing Stem Cells in Adhesion Culture

[0080] Tested herein is the animal-ingredient free ferritin-based medium substantially free of transferrin for mESC growth in 2D adherent rather than 3D suspension culture (as the latter was demonstrated in Example 2). Similar testing steps are followed as in Example 1. The mESCs are successfully cultured adherently in cell culture flasks using the animal-ingredient free ferritin-based medium containing ferritin in lieu of transferrin. The mESCs obtained at passage 12 are characterized, including viability, doubling time, morphology, pluripotency, and / or differentiation induced by removing Leukemia Inhibitory Factor. These results demonstrate ferritin is35105616786.2Atorney Docket No.: 116673-856389 able to replace transferrin in a standard serum-free, KSR-free, or animal-ingredient free medium and that the animal-ingredient free ferritin-based medium can be used successfully as a cell culture medium for growing mESCs adherently.Example 4. Animal-Ingredient Free Ferritin-Based Medium for Culturing Other Cell Types in 3D Suspension

[0081] Tested herein are different stem cell lines, including but not limited to, human induced pluripotent stem cells. The tested media include the standard KSR medium and the animal-ingredient free ferritin-based medium prepared in Example 1 , wherein ferritin substantially replaces the transferrin comprising standard KSR medium grown in 3D suspension in an ABLE vessel as in Example 2. Similar growing and testing steps are followed as in Example 2. The cell lines are successfully cultured in the suspension bioreactor in the animal-ingredient free ferritin-based medium containing ferritin in lieu of transferrin. The obtained cells at passage 12 are characterized, including viability, doubling time, morphology, and pluripotency. These results demonstrate that ferritin is able to replace transferrin in a standard animalingredient free medium.Example 5. Other Culture Media

[0082] Tested herein are different ferritin-based culture media, as detailed in Example 2 and throughout the present disclosure. Different cell lines, including but not limited to, mESCs, human induced pluripotent stem cells are cultured in these ferritin-based culture media, wherein ferritin substantially replaces transferrin. Similar testing schemes are used as in Example 1 to grow and evaluate the obtained cells.Example 6. Industrial Scale Production

[0083] Tested herein are developments and productions of ferritin-based culture media from scratch. The ferritin-based cell culture medium is produced by combining one or more ingredients selected from the group consisting of albumins or albumin substitutes, one or more amino acids, one or more vitamins, one or more ferritins, one or more antioxidants, one or more insulins or insulin substitutes, one or more inorganic36105616786.2Attorney Docket No.: 116673-856389 salts, and one or more trace elements. For example, the method comprises: (i) obtaining or having obtained ingredients of albumins or albumin substitutes, one or more amino acids, one or more vitamins, one or more antioxidants, one or more insulins or insulin substitutes, one or more inorganic salts, and one or more trace elements.; (ii) obtaining or having obtained a ferritin in a particle form; (iii) preparing the ferritin-based cell culture medium by combining the ingredients and ferritin in a gel or liquid carrier; and (iv) storing such ferritin-based cell culture medium or using it in cell culture.

[0084] Tested herein are productions of different ferritin-based culture media, optionally made from scratch as supra and / or starting with commercially available and transferrin containing cell culture media. For all productions developed, the volume of product is to be scaled up to at least 200 L, and then to 2 kL, 10 kL, move to 20 kL and 30 kL. Due to the low cost of plant-derived ferritin, such scaled up production is feasible and cost effective, making the mass production of ferritin-based cell culture medium practically viable. Various industrial scale products are to be formulated, including but not limiting to, ferritin-based cell culture media, cultivated meat compositions and products, and / or the biomass compositions or products presented herewith.Example 7. Growth Rate of Cells in Ferritin Based Culture Medium

[0085] Relative growth of cells cultured in three different media conditions, animal- free medium with ferritin, animal-free medium without transferrin or ferritin, and animal- free medium with recombinant transferrin, were tested over 47-58 passages. The control condition, recombinant transferrin in animal-free media, was set to 100% and used as a baseline for comparison. As shown in FIG. 1+2, the cells cultured in the animal-free medium with ferritin exhibited similar growth as cells cultured in animal- free medium with recombinant transferrin.Example 8. Cultivated Meat and Pet Food

[0086] A pet food “Mouse Mousse” was generated by harvesting all contents from a bioreactor, following proliferation and spontaneous differentiation. The harvested37105616786.2Attorney Docket No.: 116673-856389 contents were used as a base ingredient to which ~1% plant starches were added. This combination of 99% cultured biomass composition and 1 % plant starch was then used to produce canned wet food for cats using the standard and well-established practices in pet food manufacture. Palatability testing in animals showed general liking and consumption of the pet food product (data not shown).Example 9. Discussions and Applications

[0087] These results showed that the ferritin-based culture media were able to support both viability and long-term growth of animal cells ex vivo, making ferritin an attractive substitute for standard transferrin-based culture media. More importantly, because plant-derived ferritin is -3000 times cheaper than transferrin and can be stored at room temperature, the disclosed ferritin-based culture medium is an affordable and practical alternative media to grow cells. The ferritin used in these studies was food / feed grade, making the cells produced thereof eligible to be used in pharmaceuticals, and / or food or feed products, such as cultivated meat products.38105616786.2

Claims

Attorney Docket No.: 116673-856389WE CLAIM:

1. A cell biomass composition, wherein the composition is produced ex vivo by culturing a cell line in a culture medium comprising ferritin and substantially free of transferrin.

2. The cell biomass composition of claim 1 , wherein the ferritin is a food-grade, feedgrade, or pharmaceutical grade.

3. The cell biomass composition of claim 1 or claim 2, wherein the ferritin comprises an animal-derived ferritin, a recombinant ferritin, a plant-derived ferritin, or any combination thereof.

4. The cell biomass composition of claim 3, wherein the plant comprises a legume plant, and preferably being a Glycine max or Pisum sativum.

5. The cell biomass composition of any one of the proceeding claims, wherein the ferritin is in an amount from about 50 pg / ml to about 150 pg / ml.

6. The cell biomass composition of any one of the proceeding claims, wherein the ferritin is in an amount from about 80 pg / ml to about 120 pg / ml.

7. The cell biomass composition of any one of the proceeding claims, wherein the ferritin is in an amount of about 100 pg / ml.

8. The cell biomass composition of any one of the proceeding claims, wherein the composition is suitable for human or pet consumption.105616786.2Attorney Docket No.: 116673-8563899. The cell biomass composition of any one of the proceeding claims, wherein the composition is free of any animal ingredient or is plant based.

10. The cell biomass composition of any one of the proceeding claims, wherein the composition comprises animal ingredients.

11. The cell biomass composition of any one of the proceeding claims, wherein the composition is serum-free or serum-containing.

12. A food product, a feed product, a nutraceutical product, a dietary supplement product, a performance-enhancing product, a veterinary product, a chemical product, a scientific reagent product or a pharmaceutical product comprising the cell biomass composition of any one of the proceeding claims.

13. A cultivated meat product comprising the cell biomass composition of any one of the proceeding claims.

14. The cultivated meat product of claim 13, further comprising a functional ingredient to supplement the product’s protein content, lipid content, sugar content, fiber content, calcium content, iron content, or any combination thereof.

15. The cultivated meat product of claims 13-14, further comprising a food grade or feed grade flavorant, a food grade or feed grade colorant, a food grade or feed grade preservative, a food grade or feed grade antioxidant, or any combination thereof.

16. The cultivated meat product of claim 15, wherein the food grade or feed grade flavorant comprises a chicken flavorant, a beef flavorant, a pork flavorant, a fish flavorant, a smoke meat flavorant, a mouse flavorant, or any combination thereof.40105616786.2Attorney Docket No.: 116673-85638917. The cell biomass composition of any one of claims 1-11 , wherein the composition is lab grade, food grade, feed grade, pharmaceutical grade, or chemical grade.

18. The cell biomass composition of any one of claims 1-11 , wherein the composition is used as a culture medium component, as a carrier component, as an excipient component, as a reagent, or any combination thereof.

19. The cell biomass composition of any one of claims 1-11 , wherein the composition is used as a culture medium for culturing and / or proliferating cells suitable for a cultivated meat product.

20. The cell biomass composition of claim 19, wherein the cells suitable for a cultivated meat product comprise stem cells, fibroblast cells, immortalized cells, or any combination thereof.

21. A cultivated meat composition, comprising(i) ferritin;(ii) a culture medium substantially free of transferrin; and(iii) cells, wherein the cells are capable of being cultured and / or proliferated in the culture medium and ferritin.

22. The cultivated meat composition of claim 21 , wherein the ferritin is in an amount sufficient to support the cell line growth.41105616786.2Attorney Docket No.: 116673-85638923. The cultivated meat composition of any one of claims 21-22, wherein the ferritin is in an amount from about 50 pg / ml to about 150 pg / ml.

24. The cultivated meat composition of any one of claims 21-23, wherein the ferritin is in an amount from about 80 pg / ml to about 120 pg / ml.

25. The cultivated meat composition of any one of claims 21-24, wherein the ferritin is in an amount of about 100 pg / ml.

26. The cultivated meat composition of any one of claims 21-25, wherein the ferritin comprises an animal-derived ferritin, a recombinant ferritin, a plant-derived ferritin, or any combination thereof.

27. The cultivated meat composition of any one of claims 21-26, wherein the plant comprises a legume plant, and preferably being a Glycine max or Pisum sativum.

28. The cultivated meat composition of claims 21-27, wherein the cell is selected from stem cell, a fibroblast cell, an immortalized cell line, and any combination thereof.

29. The cultivated meat compositions of claim 28, wherein the stem cell comprises an embryonic stem cell line, an induced pluripotent stem cell line, or any combination thereof.

30. The cultivated meat composition of claim 28, wherein the immortalized cell line comprises a myoblast cell line, an adipocyte cell line, or any combination thereof.42105616786.2Attorney Docket No.: 116673-85638931. The cultivated meat composition of any one of claims 21-30, wherein the ferritin is suitable for human or pet consumption.

32. A food product, a feed product, a nutraceutical product, a dietary supplement product, a performance-enhancing product, a veterinary product, a pharmaceutical product, or a reagent to produce a chemical product comprising the cultivated meat composition of any one of claims 21-31 .

33. The cultivated meat composition of any one of claims 21-32, further comprising a functional ingredient to supplement the composition’s protein content, lipid content, sugar content, fiber content, calcium content, iron content, or any combination thereof.

34. The cultivated meat composition of any one of claims 21-33, further comprising a food grade or feed grade flavorant, a food grade or feed grade colorant, a food grade or feed grade preservative, a food grade or feed grade antioxidant, or any combination thereof.

35. The cultivated meat composition of claim 34, wherein the food or feed-grade flavorant comprises a chicken flavorant, a beef flavorant, a pork flavorant, a fish flavorant, a smoke meat flavorant, a mouse flavorant or any combination thereof.

36. A method of producing a cultivated meat composition, comprising:(i) obtaining or having obtained a cell line;(ii) culturing the cell line in a transferrin-based culture medium;(iii) transferring the cultured cells into a mixed culture medium comprising the transferrin-based culture medium and a ferritin-based culture medium, wherein the transferrin-based culture medium accounts for about 30% (v / v) to about 50% (v / v) of the mixed culture medium;43105616786.2Attorney Docket No.: 116673-856389(iv) passaging the cultured cells of step (iii) into the ferritin-based culture medium; and(v) continuing passaging the cells in the ferritin-based culture medium;(vi) harvesting the cultured cells alone, the cultured cells together with the spent media, or the biomass inside the bioreactor, and thereby producing the cultivated meat composition; wherein the transferrin-based culture medium is substantially free of ferritin; and wherein the ferritin-based culture medium is substantially free of transferrin.

37. The method of claim 36, wherein the mixed medium comprising about 50% (v / v) of the transferrin-based culture medium and about 50% (v / v) of the ferritin-based culture medium.

38. The method of claims 36-37, wherein the transferrin-based culture medium is a commercially available culture medium or a non-commercially available culture medium.

39. The method of claims 36-38, wherein the transferrin-based culture medium is KnockOut™ Serum Replacement (KSR medium).

40. The method of any one of claims 36-39, wherein the ferritin-based culture medium comprises no component from KSR.

41. The method of any one of claims 36-40, wherein the ferritin-based medium comprises ferritin in an amount sufficient to support the cell line proliferation.44105616786.2Attorney Docket No.: 116673-85638942. The method of any one of claims 36-41, wherein the ferritin-based medium comprises ferritin in an amount from about 50 pg / ml to about 150 pg / ml.

43. The method of any one of claims 36-42, wherein the ferritin-based medium comprises ferritin is in an amount from about 80 pg / ml to about 120 pg / ml.

44. The method of any one of claims 36-43, wherein the ferritin-based medium comprises ferritin is in an amount of about 100 pg / ml.

45. The method of any one of claims 36-44, wherein the ferritin in the ferritin-based medium comprises an animal-derived ferritin, a recombinant ferritin, a plant-derived ferritin, or any combination thereof.

46. The method of claim 45, wherein the plant comprises a legume plant, and preferably being a Glycine max or Pisum sativum.

47. The method of any one of claims 36-46, wherein the ferritin in the ferritin-based medium is suitable for human or pet consumption.

48. The method of any one of claims 36-47, wherein the cell line comprises a stem cell, a fibroblast cell, an immortalized cell line, or any combination thereof.

49. The method of claim 48, wherein the stem cell comprises an embryonic stem cell, an induced pluripotent stem cell, or any combination thereof.

50. The method of claim 48, wherein the immortalized cell line comprises a myoblast cell line, an adipocyte cell line, or any combination thereof.45105616786.2Attorney Docket No.: 116673-85638951. A cell culture medium comprising ferritin and substantially free of transferrin.

52. The cell culture medium of claim 51 , wherein the ferritin is in an amount sufficient to support cell proliferation from a cell line.

53. The cell culture medium of any one of claims 51-52, wherein the ferritin is in an amount from about 50 pg / ml to about 150 pg / ml.

54. The cell culture medium of any one of claims 51-53, wherein the ferritin is in an amount from about 80 pg / ml to about 120 pg / ml.

55. The cell culture medium of any one of claims 51-54, wherein the ferritin is in an amount of about 100 pg / ml.

56. The cell culture medium of any one of claims 51-55, wherein the ferritin comprises an animal-derived ferritin, a recombinant ferritin, a plant-derived ferritin, or any combination thereof.

57. The cell culture medium of claim 54, wherein the plant comprises a legume plant, and preferably being a glycine max or Pisum sativum.

58. The cell culture medium of any one of claims 50-55, wherein the cell line comprises a stem cell line, a fibroblast cell line, a cancer cell line, an immortalized cell line or any combination thereof.46105616786.2Attorney Docket No.: 116673-85638959. The cell culture medium of claim 56, wherein the stem cell comprises an embryonic stem cell, an induced pluripotent stem cell, or any cell line combination thereof.

60. The cell culture medium of claim 56, wherein the cancer cell line comprises a Hela cell line, a CHO cell line, or any cell line combination thereof.

61. The cell culture medium of claim 56, wherein the immortalized cell line comprises a myoblast cell line, an adipocyte cell line, or any combination thereof.47105616786.2

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