Effects of Treatment with Bone Morphogenetic Protein 4 and Co-culture on Expression of Piwil2 Gene in Mouse Differentiated Embryonic Stem Cells

Document Type : Original Article

Authors

1 Anatomical Sciences Department, Medical Sciences Faculty, Tarbiat Modares University, Tehran, Iran

2 Biotechnology Department, Medical Sciences Faculty, Tarbiat Modares University, Tehran, Iran

Abstract

Background
Specific growth factors and feeder layers seem to have important roles in in vitro embryonic stem cells (ESCs) differentiation. In this study,the effects of bone morphogenetic protein 4 (BMP4) and mouse embryonic fibroblasts (MEFs) co-culture system on germ cell differentiation from mouse ESCs were studied.


Materials and methods
Cell suspension was prepared from one-day-old embryoid body (EB) and cultured for four days in DMEM medium containing 20% fetal bovine serum (FBS) in the following groups: simple culture (SC), simple culture with BMP4 (SCB), co-culture (CO-C) and co-culture with BMP4 (CO-CB). Expression of piwi-like homolog 2 (Piwil2), the germ cell-specific gene, was evaluated in the different study groups by using quantitative real time polymerase chain reaction (RT-PCR). Testis was used as a positive control.


Results
The maximum and minimum Piwil2 expression was observed in SC and SCB groups, respectively. A significant difference was observed in Piwil2 expression between SCB and other study groups (p <0.05).


Conclusion
The findings of this study showed that neither the addition of BMP4 in culture medium nor the use of MEFs as a feeder layer have a positive effect on late germ cell induction from mouse ESCs.

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