Dervation Of Mouse Induced Pluripotent Stem Cells With High Potency

One dozen years ago,Takahashi and Yamanaka reported on the induction pluripotent stem cells?iPSC?from adult fibroblast cell by integrating pluripotency-associated factors Oct4,Sox2,Klf4 and c-Myc.With more and depth research on iPSCs,the major laboratories have focus on developing different methods to culture induced pluripotent stem cells.In this experiment,mouse i PSCs derived from somatic cells were reprogrammed into a novel advanced stem cells?iPSC-ASCs?with ABCL medium^[1]?Activin A,BMP4,CHIR99021,and mLif?.The derivation and characterization of iPSC-ASCs were summarized in below.1.The mouse embryonic fibroblasts cells?MEF?were transformed into iPS cells when introduced with expressed Oct4,Sox2,Klf4,c-Myc,Rarg and Lrh1.The RT-qPCR analysis of iPSCs revealed expression of Oct4,Sox2,Nanog,and AP staining was positive.2.Depend on the different morphology,iPSC were divided into flat?F-iPSCs?and round?R-iPSCs?groups.The immunofluorescence stainning of Cdx1 and c-Myc increased in both iPSC-ASCs compared to iPSCs.3.The RT-qPCR results showed the expression of pluripotent genes Oct4,Sox2 and Nanog in both i PSCs and iPSC-ASCs was significant higher than MEF.The expression of specification lineage marker T,K18 and Gata4 were increased in F-iPSC-ASCs group compare to F-iPSCs,whereas do not found same result in R-iPSC-ASCs group.4.To assess the pluripotency of iPSC-ASCs,we carried on production of chimeras test.The results showed that both group of F-iPSC-ASCs and R-iPSC-ASCs contributed to the fetus,placenta,yolk sac and germline transmission at E13.5.5.In final step,we tested the lineage differentiation in iPSC-ASCs.When iPSCs and iPSC-ASCs cultured in N2B27 basic medium for 6 days,we found pluripotent marker Oct4,Nanog and Sox2and lineage specific marker T,K18 and Gata4 were higher expression in iPSC-ASCs than iPSCs.In here,we showed the iPSC-ASCs was a novel and distinct self-renewing type of pluripotent stem cells,it exhibit specific features,including high express pluripotent marker,lineage marker.Importantly,iPSC-ASCs contributed to embryo,yolk sac and placenta in chimeras,revealed it is high pluripotent stem cell.