Functional Analysis Of Aszl In PGC Formation From Murine Embryonic Stem Cells

Embryonic stem cells (ESCs) have the capacity to self-renew and differentiate into cells of all tissue types (ectoderm, mesoderm, and endoderm). The study for embryonic stem cells has become a hot area of life sciences and medical research, and has great impact on understanding the nature of human and mammalian life. Germ cells act as cellular elements to pass genetic information from one generation to the next.. In this study, by using murine embryonic stem cells (ESCs) in vitro differentiation system, we try to identify genes whose function are important during the process from ESCs to PGCs.We choose some candidate genes and established stable cell lines by overexpressing or knockdown these specific genes by retrovirus or lentivirus infection, using these stable cell lines, we made embryoid body and examined the percentage of SSEA1positive cells at D5and D9by fluorescence-activation cell sorting(FACS). In addition, we further validate the results of SSEA1staining by alkaline phosphatase activity (AP) assay. Our data revealed that higher percentage of SSEA1staining and more AP positive clones in Aszl overexpressing cells when compare to control cells. On the contrast, the stable cell line with Aszl knockdown showed fewer AP positive clones than control. Furthermore, Q-PCR results demonstrated that the expression levels of some germ cell specific genes such as Dazl, Vasa, Oct4were higher in Aszl overexpressing lines than control cells, while the expression levels of these germ cell specific genes such as Dazl, Vasa, Oct4were lower upon Aszl knockdown. Taken together, our data suggested Aszl promoted primordial germ cells formation from murine embryonic stem cells.