17 results about "Cryoprotective Agent" patented technology

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.

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 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 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 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.

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.

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 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.

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.

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.

The present disclosure provides cryopreservation formulations, cryopreservation media, and methods of cryopreserving mammalian cells or tissues. The cryopreservation preparation and the culture medium contain a combination of cryoprotectants selected from the group consisting of: (a) glycerol and trehalose; (b) glycerol, trehalose and ascorbic acid; (c) glycerol, trehalose and taurine; and (d) glycerol, trehalose, ascorbic acid and taurine. In certain embodiments, the cryopreservation formulations and media are free of dimethyl sulfoxide.