The effects of BMP4 and BMP2 on Expression of Trophoblast-Specific Genes in Human Embryonic Stem Cells using SNU-hES3 cell line / 대한산부인과학회지

ABSTRACT

OBJECTIVE: Human embryonic stem cell derived from blastocyst randomly differentiates into multiple cell types during embryoid body development. Bone morphogenetic proteins (BMPs) are members of the transforming growth factor type beta superfamily. We have a question whether BMP2 and/or BMP4 can induce trophoblast specific genes in human ES cells using SNU hES3 cell line. METHODS: Human embryonic stem cell line (SNU hES3) was supplied by Miz Medi Hospital Seoul National University. Cultured hES cells were divided into small clumps and then allow for EB formation in differentiation medium. After EB formation, EBs were transferred onto gelatin coated dishes and given hES conditioned medium alone (control) or supplemented as following treatment for 6 days; rhBMP4 100 ng/mL; rhBMP2 100 ng/mL; BMP4 100 ng/mL +BMP2 100 ng/mL. RT PCR was performed for trophoblast specific genes. During culture, supernatant was collected and measured for estradiol (E2), progesterone, and hCG beta by enzymeimmuno assay (EIA) kit. RESULTS: BMP4 and BMP2 increase chorionic gonadotropin beta (hCG beta), glical cell missing 1 (GMC1), and CD9 as trophoblast specific gene markers confirmed by RT PCR. However, the non classical HLA class I molecule HLAG1, was not expressed in our studies. And we cannot find significant differences of the level of estradiol, hCG nd progesterone in this study. CONCLUSION: The results suggest that BMP 4 and 2 have an additive effect on induction of trophoblast related genes in SNU hES3 cell line. Although we failed to induce the differentiation of human ES cells to trophoblast, this study could provide the possibility for the differentiation of early human trophoblast cells and thus need further studies.