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

• Main Authors: Zohreh Makoolati, Mansoureh Movahedin, Mehdi Forouzandeh-Moghadam
• Format: Article
• Language: English
• Published: Royan Institute (ACECR), Tehran 2009-08-01
• Series: International Journal of Fertility and Sterility
• Subjects: embryonic stem cell; germ cell; bmp4; piwil2; co-culture
• Online Access: http://www.ijfs.ir/article_45752_9e9fd5274c8eabc8bebec6eb69e05974.pdf

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.