Parthenogenic Activation of Human Oocytes for the Production of Human Embryonic Stem Cells

Abstract

Methods of producing human stem cells are disclosed for parthenogenetically activating human oocytes by manipulation of O 2 tension, including manipulation of Ca 2+ under high O 2 tension and contacting oocytes with serine threonine kinase inhibitors under low O 2 tension, isolating inner cell masses (ICMs) from the activated oocytes, and culturing the cells of the isolated ICMs under high O 2 tension. Moreover, methods are described for the production of stems cells from activated oocytes in the absence of non-human animal products, including the use of human feeder cells/products for culturing ICM/stem cells. Stem cells produced by the disclosed methods are also described.

1 . A library of stem cells comprising autologous stem cells, wherein the stem cells are derived from parthenogenetically activated oocytes from one or more human donors. 
     
     
         2 . The library of  claim 1 , wherein each library member is identified as a full sibling, half sibling, or unrelated according to single nucleotide polymorphism (SNP) markers. 
     
     
         3 . The library of  claim 1 , wherein the oocyte donor is histocompatible with a member of the library. 
     
     
         4 . The library of  claim 1 , wherein a member of the library is genomically imprinted according to the oocyte donor origin. 
     
     
         5 . The library of  claim 1 , wherein each member is homozygous for at least one MHC allele present in a human population. 
     
     
         6 . The library of  claim 1 , wherein each member is homozygous for a different combination of MHC alleles than the other members of the library. 
     
     
         7 . The library of  claim 1 , wherein each member is at least homozygous for one or more HLA class I genes and HLA class II genes. 
     
     
         8 . The library of  claim 7 , wherein the HLA class I genes are selected from HLA A*, HLA B*, HLA and Cw* haplotype combinations. 
     
     
         9 . The library of  claim 7 , wherein the HLA class II genes are selected from HLA DRB1*, DRB3*, DRB4*, DRB5*, DQA1*, and DQB1* haplotype combinations. 
     
     
         10 . The library of  claim 1 , wherein each member (i) will proliferate in an in vitro culture for over one year, (ii) maintains the potential to differentiate to derivatives of one or all of endoderm, mesoderm, and ectoderm tissues throughout the culture, and (iii) is inhibited from differentiation when cultured on a fibroblast feeder layer. 
     
     
         11 . The library of  claim 10 , wherein each member maintains a karyotype in which the chromosomes are euploid and not altered through prolonged culture. 
     
     
         12 . The library of  claim 1 , wherein each member can differentiate into to ectoderm, mesoderm, and endoderm germ line cells.