Abstract
Germ cells are responsible for the propagation of live animals from generation to generation, but to surprise, a steep increase in infertile problems among livestock poses great threat for economic development of human race. An alternative and robust approach is essential to combat these ailments. Here, we demonstrate that goat putative embryonic stem cells (ESCs) were successfully in vitro differentiated into primordial germ cells and oocyte-like cells using bone morphogenetic protein-4 (BMP-4) and trans-retinoic acid (RA). Oocyte-like cells having distinct zonapellucida recruited adjacent somatic cells in differentiating culture to form cumulus-oocyte complexes (COCs). The putative COCs were found to express the zonapellucida specific (ZP1 and ZP2) and oocyte-specific markers. Primordial germ cell-specific markers VASA, DAZL, STELLA, and PUM1 were detected at protein and mRNA level. In addition to that, the surface architecture of these putative COCs was thoroughly visualized by the scanning electron microscope. The putative COCs were further parthenogenetically activated to develop into healthy morula, blastocysts and hatched blastocyst stage like embryos. Our findings may contribute to the fundamental understanding of mammalian germ cell biology and may provide clinical insights regarding infertility ailments.
Highlights
Germ cells are responsible for the propagation of live animals from generation to generation, but to surprise, a steep increase in infertile problems among livestock poses great threat for economic development of human race
The derivation of germ cells or oocytes from embryonic stem cells (ESCs) is a model for studying the molecular basis of the development of germ cells and may one day promote the technology of somatic cell nuclear transfer and infertility treatments
ESCs growing in LIF free DMEM with 20% fetal bovine serum (FBS), 1 μM retinoic acid and 100 ng/ml 52 bone morphogenetic protein-4 (BMP-4)
Summary
Germ cells are responsible for the propagation of live animals from generation to generation, but to surprise, a steep increase in infertile problems among livestock poses great threat for economic development of human race. We demonstrate that goat putative embryonic stem cells (ESCs) were successfully in vitro differentiated into primordial germ cells and oocyte-like cells using bone morphogenetic protein-4 (BMP-4) and trans-retinoic acid (RA). Embryonic stem cells (ESCs) are emanated from blastocyst stage embryos in mammals with unlimited proliferative capacity and multi lineages differentiation ability under suitable culture onditions[5,6,7,8]. These cells hold a wide prospective to bestead as in vitro model system to extrapolate the mechanism of gametogenesis and a cell based method of inducing mature gametes. The derivation of germ cells or oocytes from ESCs is a model for studying the molecular basis of the development of germ cells and may one day promote the technology of somatic cell nuclear transfer and infertility treatments
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