This disclosure relates to ice nucleation formulations for cryopreservation and stabilization of biologics, and methods of use thereof.

Disclosed is a process for storage of nucleic acids from a biological sample. The process comprises providing a biological sample comprising one or more cells containing nucleic acids therein; contacting the biological sample with a vitrification medium comprising a vitrification agent and a lysing agent to form a vitrification mixture; vitrifying the vitrification mixture to generate a storage-stable sample. In various aspects, the storage-stable sample can be stored at a temperature above cryogenic temperatures, such as at room temperature, for 20 days or more.

Human induced pluripotent stem cells (hiPSCs) possess tremendous potential for tissue regeneration and banking hiPSCs by cryopreservation for their ready availability is crucial to their widespread use. However, contemporary methods for hiPSC cryopreservation are associated with both limited cell survival and high concentration of toxic cryoprotectants and/or serum. The latter may cause spontaneous differentiation and introduce xenogeneic factors, which may compromise the quality of hiPSCs. Here, sand from nature is discovered to be capable of seeding ice above −10° C., which enables cryopreservation of hiPSCs with no serum, minimized cryoprotectant, and high cell survival. Furthermore, the cryopreserved hiPSCs retain high pluripotency and functions judged by the pluripotency marker expression, cell cycle analysis, and capability of differentiation into the three germ layers. This unique sand-mediated cryopreservation method may greatly facilitate the convenient and ready availability of high-quality hiPSCs and probably many other types of cells/tissues for the emerging cell-based translational medicine.

Methods for cryopreservation of biological samples are provided. The biological samples are sub-millimeter or millimeter scale biological materials. The biological samples are pancreatic islets and stem cell derived islets. Methods for cryopreservation of islets using cryomesh and multi-step loading and unloading of CPA cocktails are provided. Methods disclosed result in vitrified and rewarmed islets with high recovery, viability and functionality. Methods are scalable for high throughput production of large amounts of vitrified and rewarmed islets for use in therapeutic transplantation.

Provided herein are constructs of micro-aggregate multicellular, minimally polarized grafts containing Leucine-rich repeat-containing G-protein coupled Receptor (LGR) expressing cells for wound therapy applications, tissue engineering, cell therapy applications, regenerative medicine applications, medical/therapeutic applications, tissue healing applications, immune therapy applications, and tissue transplant therapy applications which preferably are associated with a delivery vector/substrate/support/scaffold for direct application.

Disclosed are an anti-freezing agent for protecting a biological tissue from being damaged during a freezing treatment and a preparation method therefor. The method for preparing the anti-freezing agent involves mixing ethylene glycol, water and dimethylsulfoxide homogeneously to form a matrix, and then slowly adding sodium polyacrylate. The prepared anti-freezing agent is coated onto biological tissue during a freezing treatment such that damage to the biological tissue is reduced.

The present invention relates to methods for preventing or inhibiting ice recrystallisation in substances (e.g. biological materials and food products) which are susceptible to ice crystal growth upon cryopreservation and/or thawing therefrom. The methods relate to the use of compositions comprising poly(proline) or a variant or derivative thereof. Also provided are kits and compositions comprising poly(proline) which can be used in the methods of the invention.

Presented herein are ready to administer (RTA) retinal pigment epithelium (RPE) cell therapy compositions for the treatment of retinal degenerative diseases and injuries. A method of formulating human RPE cells for administration to a subject directly after thawing and of formulating RPE cell therapy compositions for cryopreservation and administration of the cryopreserved composition to a subject subsequent to thawing are also presented. In another aspect, the RTA composition may be formulated as a thaw and inject (TAI) composition, whereby the composition is administered by injection subsequent to thawing.

Methods of expanding tumor infiltrating lymphocytes (TILs) from cryopreserved tumor tissue and methods of using the expanded TILs in the treatment of human diseases, including cancers, are disclosed. Preparing the tumor tissue for freezing includes fragmenting the tumor tissue, and incubating the fragments in a cryopreservation medium. In an embodiment, the fragments may be incubated in the cryo preservation medium at 2° C. to 8° C. for about 30 to about 80 minutes. Freezing the fragments may be done by flash freezing using the vapor phase of liquid nitrogen, e.g. using a dry cryoshipper. In some embodiments, compositions of cryopreserved tumor tissues are disclosed.

The present invention relates to a cryopreserved in vitro cell culture comprising human pancreatic progenitor cells that co-express pancreatic-duodenal homeobox factor-1 (PDX1) and NK6 homeobox 1 (NKX6.1) and are chromogranin negative. The present invention also relates to a method for cryopreserving an in vitro population of human pancreatic progenitor cells that co-express PDX1 and NKX6.1 and are chromogranin negative.