abstract |
Isolated embryonic stem cells are produced by first, inserting a human or mammalian cell or cell nucleus into enucleated oocytes of an animal species different from the human donor cell, to form a nuclear transfer unit. The resultant units are activated by exposure to ionomycin and DMAP, and cultured on a feeder cell layer comprising fibroblasts. Cells obtained from the cultured NT units are used to obtain isolated embryonic stem cells. The human donor cell may be an epithelial cell or lymphocyte, and the animal oocyte may be an ungulate selected from the group of bovine, ovine, porcine, equine, capine and buffalo. The embryonic stem cells may be used for producing desired differentiated cells. The cells may be manipulated by introduction, removal or modification of desired genes by homologous recombination. A desired gene may be one that encodes a therapeutic enzyme, growth factor or cytokine. The cells may be used in cell transplantation therapies and for in vitro study of cell differentiation. Specific examples of conditions wherein cell transplantation therapy may be applied are 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, cartilage defects or injuries, burns, foot ulcers, vascular disease, urinary tract disease, AIDS and cancer. |