Abstract
Several inducers have been used to differentiate embryonic stem cells (ESCs) into male germ cells but the induction process has been inefficient. To solve the problem of low efficiency of inducer for ESCs differentiation into male germ cells, all-trans retinoic acid (ATRA), Am80(the retinoic acid receptor agonist), and estradiol (E2) was used to induce ESCs to differentiate into male germ cells in vitro. ESCs were cultured in media containing ATRA, Am80, or E2 respectively which can differentiate ESCs into a germ cell lineage. In process of ATRA and Am80 induction Group, germ cell-like cells can be observed in 10 days; but have no in E2 induction Group. The marker genes of germ cell: Dazl, Stra8, C-kit, Cvh, integrinα6, and integrinβ1 all showed a significant up-regulation in the expression level. The ATRA-induction group showed high expression of C-kit and Cvh around 4 days, and integrinα6 and integrinβ1 were activated on day 10, respectively, while the E2-,Am80- induction group showed a high expression of C-kit as early as 4 days immunocytochemistry results shown that, integrinα6 and integrinβ1 could be detected in the ATRA-, Am80-, and E2-induction group, Positive clones in the ATRA group were greater in number than those in the other two groups. we conclued that ATRA, Am80, and E2 can promote the expression of the corresponding genes of germ cells, and had different effect on the differentiation of ESCs into male germ cells. ATRA was the most effective inducer of germ cell differentiation.
Highlights
Embryonic stem cells (ESCs), which derive from the inner cell mass, can differentiate into almost all types of cells because of their pluripotency
ESCs were positive for the expression of stage-specific embryonic antigen SSEA1(Fig 1C and 1D).These results show that ESCs isolated and cultured in vitro are undifferentiated
The cells changes in the E2-induced group were similar to those in the Am80-induced group.All the results shown that all-trans retinoic acid (ATRA), Am80, and E2 had different effect on the differentiation of ESCs into male germ cells
Summary
Embryonic stem cells (ESCs), which derive from the inner cell mass, can differentiate into almost all types of cells because of their pluripotency. As an ideal cell model, ESCs are often used to explore cell development and differentiation, especially for germ cell development[1]. ESCs have the potential to be induced to differentiate into male germ cells in vitro. Much research has focused on the differentiation of ESCs into male germ cells[2,3,4]. In these studies, PLOS ONE | DOI:10.1371/journal.pone.0164664. PLOS ONE | DOI:10.1371/journal.pone.0164664 October 14, 2016
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