Method of Cryopreservation of Stem Cell-Derived Retinal Pigment Epithelial Cells on Polymeric Substrate
AssigneeUniversity of Southern California
InventorsDanhong Zhu, David R. Hinton, Ashish Ahuja, Mark S. Humayun
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Abstract
Disclosed herein are methods and compositions for the cryopreservation of stem cells, such as stem-cell derived retinal pigment epithelial cells, that have been seeded onto and cultured on a substrate, such as a polymeric substrate. Such cryopreserved stem cells are useful for cell therapies, such as treatment of ocular damage or disease.
Claims20
A method of cryopreserving a monolayer of seeded cells, stem cells, stem cell derived cells, or combinations thereof on a substrate, the method comprising:
- exposing a parylene substrate seeded with a monolayer of cells to a temperature ramp-down phase with a temperature reduction rate between 10° C. per 10 seconds to 10° C. per 120 seconds to a final temperature between −20° C. and −100° C., wherein the substrate has one or more characteristics that enhance a viability of the seeded cells, the characteristics selected from the group consisting of:
- (i) a coefficient of thermal expansion of the substrate;
- (ii) a substrate elasticity parameter;
- and
- (iii) a substrate thickness,
- transferring the cell-seeded substrate to an intermediate temperature between −20° C. and −100° C. for a first period of time;
- and
- maintaining the cells at a storage temperature that is lower than the intermediate temperature for a second period of time, thereby obtaining cryopreserved cells.
A cell-seeded polymer substrate configured for implantation into a target site, the polymer substrate comprising:
- a non-porous parylene substrate;
- a monolayer of seeded cells, stem cells, stem cell derived cells, or combinations thereof on the substrate,
- wherein the substrate is configured to withstand cryopreservation at a temperature ramp-down rate between 10° C. per 10 seconds to 10° C. per 120 seconds to a final temperature between −20° C. per and −100° C., and a lower storage temperature thereafter;
- wherein the substrate has one or more characteristics that enhance a viability of the seeded cells, the characteristics selected from the group consisting of:
- (i) a coefficient of thermal expansion;
- (ii) a substrate elasticity parameter;
- and
- (iii) a substrate thickness.
Dependencies are inferred from the claim text.