Cell Preservation Method for Pluripotent Stem Cells

Abstract

The present invention provides a method for freezing a stem cell or a cell derived therefrom, the method including the steps of providing a cell suspension, performing ice nucleation on the cell suspension, and lowering the temperature of the ice nucleated cell suspension to a temperature sufficiently low to allow long term storage of the stem cell. The method is preferably used for the cryopreservation of human embryonic stem cells.

1 - 47 . (canceled) 
     
     
         48 . A method for providing a stem cell transplant in a subject, the method comprising administerin2 to the subject an effective amount of a pluripotent stem cell preparation that does not comprise an exogenous biological cryoprotectant. 
     
     
         49 . The method of  claim 48 , wherein exogenous biological cryoprotectant is serum. 
     
     
         50 . The method of  claim 48 , wherein the pluripotent stem cell preparation comprises a human embryonic stem cell. 
     
     
         51 . The method of  claim 48 , wherein the pluripotent stem cell preparation comprises a. pluripotent stem cell capable of differentiating into an hepatic cell, a renal cell, a dermal cell, a cardiovascular cell, a neural cell, a skeletal cell, a pancreatic cell or a reproductive cell. 
     
     
         52 . The method of  claim 48 , wherein the pluripotent stem cell preparation comprises a pluripotent stem cell capable of differentiating into a gut epithelial cell, a chondrocyte, an osteocyte, a cardiomyocyte- like cell, a beta-like cell, and a hair follicle cell. 
     
     
         53 . The method of  claim 48 , wherein the pluripotent stem cell preparation comprises a cell viability of up to 90%. 
     
     
         54 . The method of  claim 48 , wherein the pluripotent stem cell preparation is frozen or previously frozen. 
     
     
         55 . The method of  claim 54 , wherein the pluripotent stem cell preparation comprises a pluripotent stem cell that retains pluripotency and/or an undifferentiated phenotype upon thawing. 
     
     
         56 . The method of  claim 54 , wherein the pluripotent stem cell preparation comprises a pluripotent stem cell that exhibits non-cystic growth upon thawing. 
     
     
         57 . The method of  claim 48 , wherein the pluripotent stem cell preparation comprises a pluripotent stem cell that expresses a marker selected from the group consisting of Oct-4, TRA 1-60, TRA 1-81 SSEA-3 and SSEA-4. 
     
     
         58 . The method of  claim 48 , wherein the pluripotent stem cell preparation comprises a pluripotent stem cell that exhibits negligible retention of the marker SSEA-1. 
     
     
         59 . The method of  claim 48 , wherein the pluripotent stem cell preparation comprises a pluripotent stem cell that is capable of differentiating in vivo or in vitro into an endodermal, mesodermal or ectodermal cell. 
     
     
         60 . The method of  claim 48 , wherein the pluripotent stem cell preparation comprises a pluripotent stem cell that comprises a normal karyotype. 
     
     
         61 . The method of  claim 48 , wherein the pluripotent stem cell preparation comprises a pluripotent stem cell that exhibits a growth rate that is normal for a pluripotent stem cell. 
     
     
         62 . The method of  claim 48 , wherein the pluripotent stem cell preparation comprises a pharmaceutically acceptable carrier. 
     
     
         63 . The method of  claim 48  which is for treating a subject in need of a stem cell transplant or for treating or preventing a condition in a subject that requires a stem cell transplant. 
     
     
         64 . The method of  claim 63 , wherein the subject or the condition in the subject comprises a neurodegenerative disorder, heart disease, and/or diabetes. 
     
     
         65 . The pluripotent stem cell preparation of  claim 62 , whip h is a pharmaceutical composition for use by a subject in need of a stem cell transplant. 
     
     
         66 . The pluripotent stem cell preparation of  claim 62 , which is a pharmaceutical composition for use by a subject having a neurodegenerative disorder, heart disease and/or diabetes. 
     
     
         67 . The pluripotent stem cell preparation of  claim 62 , which comprises a human embryonic stem cell.