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 - 19 . (canceled) 
     
     
         20 . A differentiated cell derived from a stem cell obtained from a parthenogenetically activated oocyte from a human donor. 
     
     
         21 . The differentiated cell of  claim 20 , wherein the differentiated cell is histocompatible with the oocyte donor. 
     
     
         22 . The differentiated cell of  claim 20 , wherein the differentiated cell is selected from the group consisting of a neuronal cell, cardiac cell, smooth muscle cell, striated muscle cell, endothelial cell, osteoblast, oligodendrocyte, hematopoietic cell, adipose cell, stromal cell, chondrocyte, astrocyte, dendritic cell, keratinocyte, pancreatic islet, lymphoid precursor cell, mast cell, mesodermal cell, and endodermal cell. 
     
     
         23 . The differentiated cell of  claim 22 , wherein the cell expresses neurofiliment 68, NCAM, beta III-tubulin, GFAP, or a combination thereof. 
     
     
         24 . The differentiated cell of  claim 22 , wherein the cell expresses alpha-actinin, desmin, or a combination thereof. 
     
     
         25 . The differentiated cell of  claim 22 , wherein the cell expresses PECAM-1, VE-Cadherin, or a combination thereof 
     
     
         26 . The differentiated cell of  claim 22 , wherein the cell expresses alpha-fetoprotein. 
     
     
         27 . The differentiated cell of  claim 22 , wherein the cell comprises homoplasmic mitochondrial DNA (mtDNA). 
     
     
         28 - 39 . (canceled) 
     
     
         40 . A method of treating a subject in need thereof, comprising administering a cellular composition comprising differentiated cells, wherein the differentiated cells are derived from a stem cell obtained from a parthenogenetically activated oocyte from a human donor. 
     
     
         41 . The method according to  claim 40 , wherein the differentiated cell is selected from the group consisting of a neuronal cell, cardiac cell, smooth muscle cell, striated muscle cell, endothelial cell, osteoblast, oligodendrocyte, hematopoietic cell, adipose cell, stromal cell, chondrocyte, astrocyte, dendritic cell, keratinocyte, pancreatic islet, lymphoid precursor cell, mast cell, mesodermal cell, and endodermal cell. 
     
     
         42 . The method according to  claim 40 , wherein the subject presents a disease selected from the group consisting of Parkinson's disease, Huntington's disease, Alzheimer's disease, ALS, spinal cord defects or injuries, multiple sclerosis, muscular dystrophy, cystic fibrosis, liver disease, diabetes, heart disease, macular degeneration, cartilage defects or injuries, burns, foot ulcers, vascular disease, urinary tract disease, AIDS, and cancer. 
     
     
         43 . (canceled)