Functional Concentrations of BMP4 on Differentiation of Mouse Embryonic Stem Cells to Primordial Germ Cells

Abstract

Background Bone morphogenetic protein 4 (BMP4) has a significant role in primordial germ cells (PGCs) differentiation from mouse embryonic stem cell (mESC). The aim of this study is to determine the best concentration of BMP4 at a time of two days on differentiation PGCs from mESC. Materials and Methods To differentiate PGCs, embryoid bodies (EBs) from mESCs were cultured in concentrations of 0, 5 and 10 ng/ml BMP4 for two days. Germ cell markers Oct4 (Pou5f1), Stella (Dppa3) and Mvh (Ddx4) were analyzed by flow cytometry, immunocytochemistry and reverse transcriptase polymerase chain reaction (RT-PCR). Results Flow cytometry data demonstrated most Mvh-positive cells were observed only in the treated groups. Immunocytochemistry of EBs in the treated groups identified cells positive for Mvh. PCR results showed expression of Oct4 in the control group and treated groups. Stella and Mvh were expressed only in the treated groups. Conclusion Low concentrations of BMP4 during two days had an optimal effect on differentiation of PGCs from mESC.

Keywords

• bmp4
• cgr8
• optimal doses
• mvh positive cells
• pgcs