32 results about "Cryoprotective Agent" patented technology

Disclosed herein include methods, compositions, and kits suitable for use in preservation (e.g., cryopreservation) of samples. The composition (e.g., cryopreservation reagent) can comprise: one or more cryoprotective agent(s), one or more culture media component(s), one or more carbohydrate(s), and / or one or more amino acid(s). The composition can be capable of maintaining one or more cellular properties and / or cell viability of a plurality of cells in a sample for a period of time under a storage condition. Also provided are methods of sample preservation and sample analysis. The method can comprise contacting a sample with the cryopreservation reagent, thereby generating a shielded sample. The method can comprise exposing the shielded sample to a freezing storage condition, thereby generating a cryopreserved sample.

The application belongs to the technical field of agricultural planting and particularly relates to a freezing-preventing agent for preventing frost damage of apricot flowers and fruits and a preparation method thereof. The freezing-preventing agent is prepared from the following raw materials in percentage by mass: a penetration regulating component 3%, an antioxidant component 0.1%, mineral elements 0.5%, a film forming agent 1.5%, a plant growth regulator 0.01%-0.05%, and water, with the total being 100%. The penetration regulating component is at least one of an anti-freezing sugar, an amino acid and a penetration regulating enzyme. The antioxidant component is at least one of vitamin C, vitamin E, anthocyanin, flavone, superoxide dismutase, peroxidase and catalase.

Relates to a pharmaceutical composition and application thereof, in particular to a pharmaceutical composition which comprises cells and pharmaceutically acceptable auxiliary materials, the auxiliary materials comprise a diluent and a cryoprotective agent, the cells are T cells containing and / or expressing a chimeric antigen receptor and / or containing a coding sequence of the chimeric antigen receptor, and the T cells are T cells containing and / or expressing the chimeric antigen receptor and / or containing a coding sequence of the chimeric antigen receptor. An antigen binding domain of the chimeric antigen receptor comprises a mesothelin binding molecule containing an anti-mesothelin single domain antibody, the anti-mesothelin single domain antibody comprises CDR1, CDR2 and CDR3, the CDR1 comprises a sequence as shown in SEQ ID NO: 1, the CDR2 comprises a sequence as shown in SEQ ID NO: 2, and the CDR3 comprises a sequence as shown in SEQ ID NO: 3.

An object of the present invention is to provide an agent for promoting undifferentiation that dissolves a drug, as DMSO or the like does, but is capable of promoting undifferentiation without inducing cell differentiation when added to a medium. The agent for promoting undifferentiation of the present invention has an aprotic zwitterion represented by the following formula (1)wherein A is an anion selected from the group consisting of SO3−, —COO−, —OP═O(H)O−, —OP═O(CH3)O− and —OP═O(OR3)O−, R1 is an alkyl group having 1 to 8 carbon atoms and optionally containing one or two oxygen atoms in the molecular chain, R2 is an alkylene group having 3 to 5 carbon atoms, and R3 is hydrogen or an alkyl group optionally having a heteroatom in the molecular chain.

Cryoprotectant toxicity is a long-standing and limiting factor for the cryopreservation of many living systems. We now disclose methods by which this toxicity can be reduced or even eliminated by pre-exposing living cells, tissues, organs, and simple organisms to hydrogen peroxide or the equivalent and / or to non-toxic concentrations of cryoprotectants.

An object of the present invention is to provide an agent for promoting undifferentiation that dissolves a drug, as DMSO or the like does, but is capable of promoting undifferentiation without inducing cell differentiation when added to a medium. The agent for promoting undifferentiation of the present invention has an aprotic zwitterion represented by the following formula (1)wherein A is an anion selected from the group consisting of SO3−, —COO−, —OP═O(H)O−, —OP═O(CH3)O− and —OP═O(OR3)O−, R1 is an alkyl group having 1 to 8 carbon atoms and optionally containing one or two oxygen atoms in the molecular chain, R2 is an alkylene group having 3 to 5 carbon atoms, and R3 is hydrogen or an alkyl group optionally having a heteroatom in the molecular chain.

Provided herein are systems, methods, and cryoprotective solutions for reversible cryopreservation of biological specimens, whole organs, and whole organisms. Exemplary methods include loading a cryoprotective agent into the biological specimen, cooling the biological specimen to a cryogenic temperature for preservation, storing the biological specimen at a preservation temperature state to preserve the biological specimen, rewarming the biological specimen by increasing a temperature of the biological specimen above the preservation temperature state, and unloading the cryoprotective agent from the biological specimen. The cooling is performed at a first rate to reduce ice formation, substantially homogeneously to reduce propensity for cracking of the preserved biological specimen, and at a first pressure to prevent or reduce ice expansion within the preserved biological specimen. The rewarming is performed at a second rate to reduce ice formation, substantially homogeneously to reduce propensity for cracking, and at a second pressure to prevent or reduce ice expansion.

The invention discloses a snow flea antifreeze protein based on a cell penetrating peptide and application thereof, and belongs to the crossing field of genetic engineering and cell preservation technologies. The fusion protein rTAT-GS-MU obtained by fusing TAT peptide at the N end of rSfAFP-MU through a (G4S) 3 linker and a non-covalent compound of SfAFP-MU and Pep-1 are constructed, the intracellular delivery efficiency of the SfAFP-MU can be remarkably improved, the mitochondrial function of cells can be remarkably improved, and nuclear DNA damage can be reduced (the content of gamma-H2AX is reduced by 140% at most). The invention provides an important basis for developing a high-efficiency and low-toxicity broad-spectrum cell cryopreservation protective agent.

The application belongs to the technical field of super-low temperature storage of agricultural products, and discloses a method and a device for super-low temperature storage of garlic callus, wherein callus with good growth potential is subjected to pre-culture; the callus is loaded, dehydrated and vitrified by using a cryoprotective agent; the callus is warmed after being treated by the cryoprotective agent; the callus is recovered after being treated by the cryoprotective agent; and the recovered callus is inoculated into a culture medium for primary culture and propagation. The method can effectively improve the super-low temperature storage effect of garlic callus, and improve the survival rate of the callus after super-low temperature storage. The pre-culture time is 2 days, the loading medium loading time is 20 minutes, and the PVS2 vitrification solution treatment time is 15 minutes, so that the survival rate of the callus is obviously improved. The method can be applied to super-low temperature storage of garlic callus, and is simple in operation process and easy to apply to production, and can be used for efficient and safe storage of garlic germplasm resources.

The application relates to the field of biological medicine, and discloses a high-activity stable thrombin powder preparation and a preparation method thereof. The preparation is composed of spherical particles with a micron-nanometer hierarchical porous structure, and components include thrombin, a biocompatible skeleton material, a cryoprotective agent, a composite pore-forming agent and a surface amphiphilic modifier. The preparation method locks the activity through ultrasonic atomization and deep cold solidification of liquid nitrogen, and performs in-situ pore-forming through stepwise temperature control and sublimation drying. Through the synergistic effect of the hierarchical pore structure and the surface amphiphilic modification, the moisture resistance and flowability of the powder preparation are enhanced, the blood is rapidly perfused by utilizing capillary force, and the high-activity maintenance and second-level hemostasis response of the thrombin are realized.

The invention discloses an ovarian tissue low-temperature cryopreservation solution as well as a preparation method and application thereof, and relates to the technical field of biomedical materials. The invention relates to an ovarian tissue low-temperature cryopreservation solution as well as a preparation method and application thereof. Every 100 ml of the ovarian tissue low-temperature cryopreservation solution comprises 15-25 ml of dimethyl sulfoxide, 15-25 ml of ethylene glycol, 15-20 g of polysucrose, 38-46 g of sucrose, 40-45 ml of a phosphate buffer solution and 0.12-0.13 g of human serum albumin. According to the low-temperature cryopreservation solution for the ovarian tissues, the dimethyl sulfoxide and the ethylene glycol are used as permeable low-temperature protective agents to prevent the ovarian tissues from being damaged at low temperature, and the polysucrose and the sucrose are used for regulating the cell osmotic pressure to improve the cell dehydration effect, reduce the formation of ice crystals in cells and increase the viscosity of the solution; according to the low-temperature cryopreservation solution for the ovarian tissue, the solution forms an amorphous glassy state instead of ice crystals in the freezing process, the human serum albumin maintains osmotic pressure and nutrition, the tissue survival rate and the functional integrity are guaranteed, the ovarian tissue can be efficiently cryopreserved through the low-temperature cryopreservation solution for the ovarian tissue, and the follicle survival rate and the functional integrity after ovarian transplantation are effectively improved.

Disclosed herein include methods, compositions, and kits suitable for use in preservation (e.g., cryopreservation) of samples. The composition (e.g., cryopreservation reagent) can comprise: one or more cryoprotective agent(s), one or more culture media component(s), one or more carbohydrate(s), and / or one or more amino acid(s). The composition can be capable of maintaining one or more cellular properties and / or cell viability of a plurality of cells in a sample for a period of time under a storage condition. Also provided are methods of sample preservation and sample analysis. The method can comprise contacting a sample with the cryopreservation reagent, thereby generating a shielded sample. The method can comprise exposing the shielded sample to a freezing storage condition, thereby generating a cryopreserved sample.

The invention relates to the technical field of biological cell storage, in particular to a bone marrow stem cell cryopreservation method and device.The method comprises the steps that a bone marrow stem cell cryopreservation system is started, the bone marrow stem cell cryopreservation system comprises a stem cell screening unit, a cryoprotectant selecting unit, a cooling unit and a liquid nitrogen cryopreservation unit, and original bone marrow is confirmed; the stem cell screening unit is used for selecting the bone marrow to be screened with the highest cell survival rate to obtain bone marrow to be cryopreserved, a currently available cryoprotectant is selected from the cryoprotectant selection unit, the cryoprotectant is injected into the cooling unit, and then the bone marrow to be cryopreserved is obtained. The bone marrow cryopreservation unit is used for receiving to-be-cryopreserved bone marrow transmitted by the stem cell screening unit, cooling the to-be-cryopreserved bone marrow containing a cryoprotectant in the cooling unit to obtain cooled bone marrow, and the liquid nitrogen cryopreservation unit is used for cryopreserving the cooled bone marrow to complete cryopreservation of bone marrow stem cells. According to the invention, the intelligence of cryopreserved bone marrow stem cells can be improved.

The application provides application of a protein deacetylase inhibitor NAM in poultry semen ultra-low temperature cryopreservation and belongs to the technical field of poultry breeding preservation. The protein deacetylase inhibitor NAM is used as a cryoprotective agent or is combined with other cryoprotective agents, and the protein deacetylase inhibitor NAM is used as an effective component, so that the efficiency of the poultry ultra-low temperature cryopreservation can be improved. The protein deacetylase inhibitor NAM can inhibit sperm cell deacetylation modification, maintain a high level of acetylation modification level, improve the ability of sperm to respond to ultra-low temperature freezing stimulation, and improve the fertilization ability of frozen-thawed sperm.

The invention belongs to the technical field of biomedicine, and particularly relates to a nanocrystal nucleus anti-freezing agent system for low-temperature preservation of a corneal graft. The invention firstly provides a nanocrystal nucleus antifreezing agent which is composed of propylene glycol, ginkgo leaf alcohol extract, trehalose, gold nanoparticles and glycerin, and the nanocrystal nucleus antifreezing agent can inhibit the anti-vitrification of corneal graft, reduce the formation of ice crystals, reduce the damage to cells and tissues to the greatest extent, and improve the anti-vitrification performance of corneal graft. And the survival rate of the corneal graft after cryopreservation is improved. Therefore, the corneal graft is placed in the nanocrystal nucleus anti-freezing agent provided by the technical scheme, and the low-temperature preservation of the corneal graft can be simply and conveniently realized only in an environment of-70 DEG C to-80 DEG C.

The invention discloses oriented engineering modification and application of a snow flea antifreeze protein, and belongs to the technical field of gene engineering. According to the invention, the rSfAFP is subjected to T16G, A19G, S22G and other multi-site mutations through a genetic engineering means, and the mutant rSfAFP-MU with high anti-freezing activity is constructed and obtained. The heat stagnation activity of the mutant is increased to 5 DEG C from 2.12 DEG C of a wild type, when the mutant is used as a cryoprotectant, the recovery rate of cells can be increased by 4.3%-10.76% compared with that of the wild type, the antifreeze mechanism of the antifreeze protein of the snow flea is clarified through site-specific mutagenesis and molecular dynamics simulation, and an important basis is provided for activity modification and structural function research of the antifreeze protein.

Disclosed herein as systems, devices and methods for introducing cryoprotective agents into cells or membrane-bound cell fragments using magnetoporation, and methods of using same to prepare cells or membrane-bound cell fragments for lyophilization or cryopreservation. The cells can be blood cells, such as red blood cells or platelets.

The invention belongs to the field of cryopreservation agents, and particularly relates to a design method of antifreeze polypeptide, a polypeptide-based cryopreservation protective agent and a preparation method and application of the polypeptide-based cryopreservation protective agent. The polypeptide-based cryopreservation protective agent comprises antifreeze polypeptide, small molecules and a buffer solution, and the negative charge density of the antifreeze polypeptide is 5-25%. The cryopreservation protective agent disclosed by the invention has good biocompatibility. Wherein small molecules are used as a natural osmotic pressure regulator, can enter cells to regulate osmotic pressure balance, and can inhibit formation and growth of ice crystals in the cells; the antifreeze polypeptide is simple in structure, so that the problem of immunogenicity of natural antifreeze protein is solved. The synergistic effect of the two can effectively inhibit the growth and recrystallization of ice crystals, form a round and fine ice crystal morphology, protect cells from mechanical damage during cryopreservation, and improve the survival rate and function of frozen cells. As a preferable form of the invention, the erythrocytes are preserved in liquid nitrogen by using the cryopreservation protective agent, the survival rate of the recovered erythrocytes reaches 93%, and the morphology and oxygen carrying functions of the erythrocytes are retained.

The novel cryopreservation protective agent comprises the following components: an ice crystal inhibitor, a natural deep eutectic solvent, an impermeable protective agent, a cell membrane stabilizer, an antioxidant, an isoosmotic adjusting agent, a buffer system and a solvent. The preparation method of the novel cryopreservation protective agent comprises the following steps: S1, preparing an environment and a solvent; s2, preparing a natural deep eutectic solvent, accurately weighing choline and L-proline according to a molar ratio of 1: 1, adding 20% of water for injection, and stirring at 60 DEG C until a clear, transparent and uniform liquid natural deep eutectic solvent is formed; s3, dissolving and mixing; s4, adjusting the pH value; s5, degerming and sub-packaging; and S6, quality control. The invention relates to a clinical-grade (elution-free) human cell cryopreservation protective agent which is completely free of DMSO (dimethylsulfoxide) and free of animal-derived components. After resuscitation, cleaning is not needed, the liquid can be directly infused back to the human body, the clinical operation process is greatly simplified, and the safety is improved.

Provided herein are systems, methods, and cryoprotective solutions for reversible cryopreservation of biological specimens, whole organs, and whole organisms. Exemplary methods include loading a cryoprotective agent into the biological specimen, cooling the biological specimen to a cryogenic temperature for preservation, storing the biological specimen at a preservation temperature state to preserve the biological specimen, rewarming the biological specimen by increasing a temperature of the biological specimen above the preservation temperature state, and unloading the cryoprotective agent from the biological specimen. The cooling is performed at a first rate to reduce ice formation, substantially homogeneously to reduce propensity for cracking of the preserved biological specimen, and at a first pressure to prevent or reduce ice expansion within the preserved biological specimen. The rewarming is performed at a second rate to reduce ice formation, substantially homogeneously to reduce propensity for cracking, and at a second pressure to prevent or reduce ice expansion.

The application provides a polyamphoteric electrolyte cell cryoprotective agent and a preparation method and application of a polyamphoteric electrolyte cell cryopreservation solution, relates to the fields of biomedical materials technology and cell cryopreservation, and aims to solve the problems of low cell recovery rate and short low-temperature storage time of cells in the existing cell cryopreservation technology. The polyamphoteric electrolyte cell cryoprotective agent comprises polylysine, polyacrylic acid and graphene oxide connected through a chemical bond, the mass ratio of the polyacrylic acid and the polylysine is 10:1-2:1, and the mass ratio of the total mass of the polyacrylic acid and the polylysine to the mass of the graphene oxide is 1:2-10:1. The polyamphoteric electrolyte cell cryoprotective agent can not only reduce the osmotic damage of cells in the freezing and thawing process by adjusting the osmotic pressure, but also improve the freezing efficiency and inhibit the formation and growth of ice crystals. The protective agent can be used in the fields of stem cell cryopreservation, immune cell cryopreservation and the like due to the excellent cell protection performance and good stability.

The present application relates to a kind of fecal bacteria cryopreservation and thawing method and cryoprotectant.The concentration of permeable protective agent in the fecal bacteria cryoprotectant is 350-400mg / ml, the concentration of impermeable protective agent is 110-130mg / ml, and the concentration of two-dimensional Ti3C2Tx nanosheet is 0.2-0.6mg / ml.The present application also provides a method for cryopreservation and thawing of fecal bacteria using the cryoprotectant, comprising the following steps: (1) preparing fecal bacteria; (2) preparing alginate hydrogel loaded with fecal bacteria; (3) cryopreservation of fecal bacteria; (4) thawing.The cryoprotectant of the present application contains two-dimensional Ti3C2Tx x nanosheet, so it has passive ice inhibition and photothermal active ice inhibition function.During the cryopreservation process, two-dimensional Ti3C2Tx x nanosheet has a significant inhibitory effect on the formation and growth of ice crystals.During the thawing process, two-dimensional Ti3C2Tx x nanosheet fully exerts its own photothermal effect, absorbs the energy of near-infrared laser and converts it into heat, acts as a high-efficiency space heat source, improves the uniformity of internal rewarming of the sample, and reduces the freezing damage caused by recrystallization and devitrification.

Provided herein are systems, methods, and cryoprotective solutions for reversible cryopreservation of biological specimens, whole organs, and whole organisms. Exemplary methods include loading a cryoprotective agent into the biological specimen, cooling the biological specimen to a cryogenic temperature for preservation, storing the biological specimen at a preservation temperature state to preserve the biological specimen, rewarming the biological specimen by increasing a temperature of the biological specimen above the preservation temperature state, and unloading the cryoprotective agent from the biological specimen. The cooling is performed at a first rate to reduce ice formation, substantially homogeneously to reduce propensity for cracking of the preserved biological specimen, and at a first pressure to prevent or reduce ice expansion within the preserved biological specimen. The rewarming is performed at a second rate to reduce ice formation, substantially homogeneously to reduce propensity for cracking, and at a second pressure to prevent or reduce ice expansion.

The application discloses a system and a method for thawing and recovering biomaterials, wherein the system comprises a carrier, a dish container and a chip. The carrier is provided with a groove near the front end, and the groove is filled with biomaterials to be thawed and a cryoprotective agent. The dish container is filled with an oil-phase thawing solution, and the groove of the carrier is immersed in the oil-phase thawing solution, so that the oil-phase thawing solution covers the groove of the carrier and thaws and recovers the biomaterials. The chip is packaged with chemicals containing the cryoprotective agent and / or a basic culture solution. The carrier is removed from the oil-phase thawing solution, and the chip covers the carrier. The chip is moved relative to the carrier, and the chemicals fully cover and contact the groove to remove the cryoprotective agent in the groove. The application can effectively standardize and automate the whole thawing and recovering process, realize effective quality control, increase processing efficiency, and reduce personnel training requirements.

The application belongs to the field of cell cryopreservation, and particularly relates to a cryoprotective agent, a preparation method and application thereof, and a T lymphocyte cryopreservation and recovery method. The cryoprotective agent comprises an anti-freezing agent and a protective solution; the anti-freezing agent is a mixture of a zwitterionic magnetic nanoparticle and betaine. The zwitterionic magnetic nanoparticle comprises a core layer of ferroferric oxide and a shell layer of a zwitterionic polymer; preferably, the zwitterionic polymer is one of poly (methacrylic acid sulfobetaine), poly (2- (methacryloyloxy) ethyl) phosphorylcholine or poly (methacrylic acid carboxybetaine); the application uses the cryoprotective agent to preserve T lymphocytes in liquid nitrogen, and it is found that the survival rate of the T lymphocytes after recovery is more than 80%, and the T lymphocytes retain immune functions such as proliferation, differentiation and tumor killing, and the effect is better than that of a traditional 10% DMSO cryoprotective agent.

The invention relates to a novel preservation scheme taking propylene glycol as a low-temperature protective agent, and provides a low-temperature preservative and a preservation method for biological large tissues. The preservative takes propylene glycol as a main low-temperature protective agent, and comprises fetal calf serum and a culture medium, the pH value is adjusted to be 7.30 to 7.45. The preservation method comprises the following steps: carrying out gradient perfusion on tissues by using the preservation agent, sequentially carrying out balanced cooling in an environment of-15 DEG C and profound hypothermia transition in an environment of-80 DEG C, and transferring the tissues into liquid nitrogen for long-term preservation. According to the scheme, the characteristic that the freezing point of a propylene glycol aqueous solution mixture is reduced is utilized, a specific formula and a step-by-step cooling procedure are combined, pre-cooling is conducted at the temperature of 0-15 DEG C in a non-freezing state, ice crystal damage is effectively reduced, the activity and structural integrity of large biological tissues are better maintained, and the method is suitable for low-temperature preservation of various in-vitro biological tissues.

Provided is a composition comprising (i) a nano- or microparticle formulation of a therapeutically active agent which is suspended in a liquid phase, and (ii) at least one cryoprotective additive selected from C3-C5 alkanes substituted by one or two hydroxy groups which stabilizes the particle formulation. Further aspects relate to a solid composition which can be obtained by freezing the stabilized composition, and to processes for the preparation of the compositions in accordance with the invention.

This invention provides a cell cryopreservation method that avoids or improves upon the use of DMSO as a cryoprotective agent. [Solution] Cell material containing living cells is preserved during a cryopreservation protocol by combining the cell material with a cryoprotective formulation / medium / solution containing an effective amount of trehalose (in the absence of DMSO and / or any other added cryoprotective agents). That is, the cryopreservation protocol does not contain any cryoprotective agents other than trehalose, and the cryopreservation protocol includes exposing the cell material to a cryoprotective formulation containing an effective amount of trehalose to act as a cryoprotective agent, cooling the cell material to a predetermined temperature below -20°C at a cooling rate in the range of -3°C / min to -50°C / min, and obtaining warmed cryopreserved cell material.

This disclosure describes compositions and methods related to cryoprotection of biomaterial. Generally, the cryoprotective composition includes a cryoprotective agent and magnetic nanoparticles effective for thawing a cryopreserved specimen comprising biomaterial with minimal biomaterial damage. In some embodiments, the composition is effective for thawing a cryopreserved specimen having a minimum dimension of 0.1 mm. Generally, the method includes obtaining a biomaterial cryopreserved with a cryoprotective composition as summarized above, then subjecting the cryopreserved biomaterial to electromagnetic energy of an intensity sufficient to excite the magnetic nanoparticles and thaw the biomaterial.

Hydrogel beads having tunable rates of loading and unloading of cryoprotective agents are provided herein. Such hydrogel beads can be dispersed throughout a cell suspension to enable loading and unloading of cryoprotective agents from cells in a gradual and distributed manner that protects the cells from osmotic damage. Lymphocyte viability after cryopreservation is significantly greater when cryoprotective agents are loaded and unloaded using hydrogel beads compared to conventional media exchange methods.