The present disclosure provides for cryopreservation compositions and methods for cryopreserving biological materials using red blood cells (RBCs) to improve cell viability. RBCs loaded on the surface with target specific antibodies will allow modified RBCs in close proximity to target cells and will provide the benefits of RBC properties to target cryopreserving cell population. Furthermore. target specific antibody conjugated RBCs will be loaded with cryoprotectant molecules such as trehalose (type of sugar molecules) alone or in combination with free radical scavengers like catalases. glutathione peroxidase. superoxide dismutase (SOD), -tocopherol (Vit. E). ascorbic acid (Vit. C). carotene (Vit. A), selenium to further improve the recovery of cells post cryopreservation.

Provided is a preservation solution for low-temperature preservation of adipose-derived mesenchymal stem cells. The preservation solution contains a compound electrolyte injection, a glucose and sodium chloride injection, vitamins, and amino acids. The preservation solution can be used for preserving adipose-derived mesenchymal stem cells at a low temperature, so as to effectively maintain the viability of adipose-derived mesenchymal stem cells, and meet clinical requirements.

The present disclosure relates to a method for preserving free cells, such as blood cells, by freezing in the presence of a nontoxic cryoprotectant, i.e., one that can be administered to a living human or animal organism. The present disclosure thus relates to methods, compositions and kits for freezing and storing non-adherent cells, such as red blood cells, for extended periods of time while preventing the lesions that can occur during the storage thereof and, in the case of red blood cells, while preserving deformability and increasing survival.

A method for establishing a seed cell bank of mesenchymal stem cells (MSC), which belongs to the technical field of stem cell culture. The method includes: mixing a plurality of P2 generation (Pn1 generation) MSCs from different donor sources for culturing.

Disclosed herein are frozen formulations of apoptotic mononuclear enriched cell populations, methods of making the frozen formulations, and methods of using the frozen formulations following thawing of formulations. Frozen formulations further include an isotonic crystalloid solution and a cryoprotectant. The thawed formulations of apoptotic mononuclear enriched cells maintain the biological activity observed in fresh cells prior to freezing.

Provided are a cold storage method and a cold storage liquid that are suitable for non-freezing refrigerated storage of human or animal stem cells such as iPS cells or embryos in a non-frozen state. According to an embodiment, the non-freezing refrigerated storage liquid is: a mixture liquid (HTM, HTMx2c13), in which MEM-alpha (Minimum Essential Medium Eagle (MEM) Alpha Modification) is mixed with a UW solution (UW_sln; BELZER UW (registered trademark) COLD STORAGE SOLUTION), or a modified UW solution (polymer component being changed to PVA), are mixed in a ratio of about 1/1 to 1/2; or a storage liquid having a composition equivalent to the mixture liquid, especially in respect of concentrations of potassium ions and sodium ions. And, arousal period(s) of maintaining a temperature of 30 C. to 37 C. is inserted at each of two to five days, into a non-freezing refrigerated storage period, which is made at 2 C. to 8 C. for 2 to 10 days or 4 to 20 days.

It is to provide a liquid for cryopreserving a cell that can effectively suppress cell death caused by freezing and thawing of a mammalian cell and that can effectively increase the proliferation ability of the mammalian cell after freezing and thawing, and a method for cryopreserving a mammalian cell using the liquid for cryopreserving a cell. A liquid comprising 2.5 to 8.75 (v/v) % propylene glycol is used as a liquid for cryopreserving a mammalian cell.

A method for in vitro expansion of cryopreserved cord blood-derived regulatory T cells (Tregs) with a high recovery rate is provided, including the following steps: recovering the cryopreserved cord blood-derived Tregs after first expansion, and resuspending the cells with a serum-free medium; and adding a resulting suspension to an expansion culture medium, and conducting a second expansion culture, where a primary culture is conducted for 1 d to 2 d, then a subculture is conducted once every 1 d to 3 d, and a total culture time is 13 d or more. In the expansion culture medium, interleukin-2 (IL-2) is further added. The method can achieve the second recovery and expansion of cryopreserved Tregs produced after the first expansion. Tregs produced after the second recovery and expansion can have a viability of 90% or more, and can be further expanded (such as third expansion and fourth expansion).

Provided are methods and compositions for cryopreserving subpopulations of lymphocytes. In one aspect, provided herein is a method of generating a sub-population of lymphocytes from a biological sample, the method comprising cryopreserving said biological sample in a cryopreserving solvent comprising less than 2M molar concentration of an active cryopreserving moiety. In another aspect, provided herein is a cellular composition comprising an enhanced sub-population of lymphocytes, an enhanced second sub-population of lymphocytes, and a depleted third subpopulation of lymphocytes, wherein said cellular composition further comprises a cryopreserving solvent comprising less than 2M molar concentration of an active cryopreserving moiety. In another aspect, provided herein is a method for preparing an allogenic composition from a biological sample, wherein said allogenic composition comprises an enhanced sub-population of lymphocytes, the method comprising cryopreserving said biological sample in a cryopreserving solvent comprising less than 2M molar concentration of an active cryopreserving moiety.

Methods of expanding and cryopreserving a natural killer (NK) cell fraction for clinical use are provided, and particularly, methods for providing expanded, cryopreserved NK cell fractions and protocols for their use, which can be employed for applications in cell transplants and infusions for treatment of cancer and other disease.