جداسازي سلولهاي بنيادي جنين موش در مرحله ي بلاستوسيست

ISOLATION & PRODUCTION OF MOUSE EMBRYONIC STEM CELLS FROM LATE BLASTOCYST STAGE EMBRYOS

Introduction:Embryonic stem cells are pluripotent cells which are derived from early stage embryos and they have the ability to differentiate to several kinds of cells invitro. They are also used for genetic manipulation. Because of these properties, these cells are used in cell therapy and production of transgenic animals. The aim of this study was isolating and producing mouse embryonic stem cells from late blastocyst stage embryos for the first time in Iran for use in future investigations. Materials At Methods: 385 blastocyst stage embryos from pregnant NMRI mice were obtained and cultured 24-hours in DMEM medium supplemented with 10 % FBS. 2-4 days after hatching, inner cell masses formed colonies and they were recognized by their morphology and collected after tripsinization. Then, some of those cells were cultured on the primary mouse embryo fibroblast feeder layer and the others were cultured in the medium containing 0.1 mM b-Mercaptoethanol,1000 u/ml LIF and 10% FBS. The embryonic stem cells were recognized by alkaline phosphatase histochemical technique. Results: Our results showed that highly pluripotent stem cells can be derived from blastocyst stage NMRI mouse embryos. Primary mouse embryo fibroblast feeder layer couldnʹt affect the growth and differentiation of embryonic stem cells. Conclusion: Overall, these embryonic stem cells can be used in future research such as producing of transgenic animals, gene therapy, gene targeting, etc.