Effect Of Sequential Co-culture System On The Development And Quality Of Murine Embryos Fertilization In Vitro

IVF (In vitro fertilization, IVF) have studied more than 100 years of history. The techniques revealed gametes for the development, maturation, fertilization and early embryonic cell division in the mysteries of life phenomena is of great significance. In the process of in vitro fertilization, oocytes can mature spontaneously, but fertilization, cleavage and early embryo development in vitro the success rate is very low, in vitro fertilization and embryo development in vitro there retardation issue. To better simulate the oocyte and early embryo development in vivo micro-environment for the success of IVF. Researchers have done a lot of testing at home and abroad in recent years. Early mammalian embryo began in the oviductal and then enter the uterus, the ultimate in human development of the unborn child. Before the embryos were cultured in vitro study was restricted to a cell and has been co-cultured embryos to blastocyst. Although this system to enhance the development of the embryo in vitro growth rate and quality, but still not very good simulation of in vivo development environment. The purpose of this study is to better simulate the oocyte and early embryo development in vivo micro-environment, embryos for in vitro experiments provide a method.The methods of 0.25% trypsin digestion were isolated oviduct epithelial cells and endometrial fine Cellular and on cultured cells in vitro proliferation laws preliminary exploration, using trypan blue exclusion test live cells. Thaw frozen by the sequential cells were cultured in vitro system for a large number of seed cells; sperm were placed tubal fluid and T6 solution, can be observed by the two semen right mouse oocytes in vitro fertilization; will be an early IVF embryo with KSOM-AA fluid, oviductal epithelial cells, Endometrial cells were cultured and sequential training system were observed on mouse embryo development in vitro fertilization effect of the impact.The results show that:(1) The epithelium cells were isolated by 0.25% trypsinization, and the cell vigor over 90% and defrosting cells vigor over 85%. (2) The endometrial cells use 0.25% trypsin digestion, living cells of more than 90%, defrosting cells vigor more than 75%.(3) The better fluid of mice mature oocytes tubal in vitro fertilization is HTF.(4) Sequential system embryos were cultured in vitro development of the blastocyst was significantly higher than that of other co-culture group (P <0.05). Experimental results show a total of sequential training system than the total integration of cell culture systems to better simulate the in vivo environment.(5) The preliminary establishment of a mouse embryo in vitro fertilization in vitro co-culture system for sequential, for the next step in laying the foundation for research.