Isolation And Identification Of Murine Female Induced Pluripotent Stem Cells

Yamanaka and his team for the first time successfully generate Mouse induced pluripotent stem cells(Induced pluripotent stem cells, i PSCs) in 2006, due to the potential for broad application prospects i PS cells,it is rapidly become new research hotspot in the field of stem cell.The creation of high quality induced pluripotent stem cells(i PSCs) has been determined as high-grade chimeras that are competent for germline transmission and viable all-i PSC mice can be generated through tetraploid complementation. Most of the high quality i PSCs described to date are male.Scientists study pluripotent stem cell lines are few female i PSCs, including cells provided by commercial companies, including ES and i PS cells, are mostly male. Therefore, the quality of female i PS cell lines for clinical research cell research applications and the study of human pluripotent female i PSCs is particularly important. Female i PSCs, especially fully pluripotent female i PSCs, are also essential for clinical application and scientific research. Here, we show, for the first time, that a gender-mixed induction strategy could lead to a skewed sex ratio of i PSCs.The tests were divided into five groups, the number of initial cells and passages were same.After reprogramming, 0% 10%,30%,50%and100%male initiating mouse embryonic fibroblasts(MEFs) at different male ratio resulted in 10%, 30% and 50% male cells yielded 0%,19.9 ±2.8%(P<0.05), 68.2 ±5.4%(P<0.05), and 85.9 ±6.8%(P<0.05), 100%male i PSCs, respectively. However, separated male and female i PSCs showed no evident differences. Furthermore, these female i PSCs had pluripotent properties typical of embryonic stem cells(ESCs): doubling speed, a ES colony-like growth, high alkaline phosphatase activity, expression of ES cell-specific surface antigens, expression of all pluripotent genes in ES cells, pluripotent gene promoter region is maintained hypomethylation. i PS cells were cultured in vitro can spontaneously form embryoid bodies, and there is spontaneous differentiation. The i PS cells were injected into immunodeficient mice after about 4 weeks, teratoma can be formed, after slicing, we can see typical three germ layers. Chimeric mice which had germline transmission capacity can be obtained by chimera experiments by use of i PS cells. Most importantly, these fully pluripotent female i PSCs could generate viable mice by tetraploid complementation. These findings indicated that high quality female i PSCs can be derived effectively by reprogramming pure female somatic cells and suggest that clinical application of female i PSCs is feasible.