Effect Of Granulosa Cells' Mitochondria Transfer On Bovine Parthenogenetic Embryos' Developmental Potential

Mitochondria are the important semi-automatic organelle in eucaryotic cells, generating cellular energy in the form of ATP(adenosine triphosphate)by the process of oxidative phosphorylation to drive cellular various activities. Because of mitochondria don't proliferate before blastocyst hatch, so the development of pre-implantation embryos depend on oocytes'mitochondria mostly to supply energy. Whatever factor that influence the mitochondria function can reduce the embryos'development. The mitochondria number of pre- fertilization oocytes must reach a threshold so that every blastomere can get enough mitochondria, and every blastomere must have enough copies of mtDNA that can meet mtDNA duplication of the pre-implantation embryos. Embryonic development need certain ATP, if the ATP content of oocytes less than a certain threshold embryos can't develop. There has been a report that human self-granulosa cells mitochondria improved human IVF embryos development, but the effect of mitochondria transfer on parthenogenetic embryos is not reported. In this study, the culture of bovine granulosa cells,the extraction and identification of bovine granulosa cells'mitochondria and the effect of mitochondria transfer on bovine parthenogenetic embryos were discussed. The objective is to increase the development of the parthenogenetic embryos and promote the isolation of embryonic stem cells from parthenogenetic embryos and other embryos.1,The bovine granulosa cells cultured at 38℃,5% CO2,100% humidity in high glucose DMEM+10%(V/V) NBS assumed stars when they just adhibited and didn't contact each other, the most cytoplasm assumed triangles, a tiny minority of cytoplasm assumed shuttles. When they contacted each other they assumed paving stone, the boundary between cells weren't clear. The proliferation of the cells became weaker as the culture passages increased. The apoptosis rate of passage 1,3,6 granulosa cells were respective 11.45%,16.63% and 39.23%.2,Using low osmotic solution(RSB) to swell the bovine granulosa cells for 8 min, broke cells in homogenizer, added high osmotic solution(2.5×MS) to make the solution isotonic, then extract mitochondria by differential centrifugation. The outer membrane and cristae of...