METHODS FOR PRODUCTION OF ICSI AND SCNT DERIVED MACAQUE STEM CELLS

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. For this protocol, macaques will be hormonally stimulated with super-ovulatory drugs to produce oocytes that will be collected via laparotomy. These oocytes will be fertilized in the laboratory using fresh or frozen sperm collected by rectal probe electro-ejaculation. The collected gametes will be used for fertilization in the laboratory by intracytoplasmic sperm injection and the embryos cultured to the blastocyst stage. Once reaching this stage, several methods will be employed to attempt to derive embryonic stem cells. In addition, some oocytes may undergo somatic cell nuclear transfer to form cloned embryos. These embryos will also be cultured to the blastocyst stage in order to derive stem cells. Long term, we anticipate that this research will enable us to optimize methods for the derivation of macaque embryonic stem cells from both fertilized and SCNT embryos. However, for this pilot project, our goal is to simply obtain data on cyno follicular stimulation and determine whether or not cynos may be an ideal animal model for future work. Any established cell lines will undergo rigorous testing to determine their pluripotency and normalcy. While human embryonic stem cells may have incredible therapeutic value for treating a wide range of disorders such as Parkinson's diabetes, autism, heart disease, Alzheimer's etc., rigorous testing in animal models is necessary before any such trials can begin. Macaques provide a valuable non-human primate model in which to determine the safety and efficacy of immune-matched stem cells.