Department of Anatomical Sciences, Medical Sciences Faculty, Tarbiat Modares University, Tehran, Iran.
In Vitro Cell Dev Biol Anim. 2011 Jun;47(5-6):391-8. doi: 10.1007/s11626-011-9404-9. Epub 2011 Apr 27.
Presence of specific growth factors and feeder layers are thought to be important for in vitro embryonic stem cell (ESCs) differentiation. In this study, the effect of bone morphogenetic protein 4 (BMP4) and mouse embryonic fibroblasts (MEFs) co-culture system on germ cell differentiation from mouse ESCs was evaluated. One-day-old embryoid body was cultured for 4 d in simple culture systems or on top of the MEFs, both in the presence or absence of BMP4. Data showed significant higher viability percent and proliferation rate in simple culture media compared to co-culture systems. Analysis of gene expression indicated that the germ cell-specific genes (VASA and Stra8) were expressed in a significant higher ratio in BMP4-treated cells in simple culture system. Also, the results of immunocytochemistry in simple culture systems showed that the mean percentage of immunostaining cells of VASA, the primordial germ cell (PGC) marker, was increased significantly in BMP4-treated cells compared with BMP4-free group. Meanwhile, CDH1, the late premiotic germ cell marker, showed no significant difference between these two groups. The results suggest that BMP4 is an efficient inducer in PGC derivation from mouse ESC. However, the employment of MEFs as feeder has no apparent effect on PGC derivation.
存在特定的生长因子和饲养层被认为对体外胚胎干细胞（ESCs）分化很重要。在这项研究中，评估了骨形态发生蛋白 4（BMP4）和小鼠胚胎成纤维细胞（MEFs）共培养系统对小鼠 ESCs 生殖细胞分化的影响。将 1 天大的胚体在简单培养系统中或在 MEFs 上培养 4 天，同时存在或不存在 BMP4。数据显示，与共培养系统相比，简单培养培养基中的细胞活力百分比和增殖率显著更高。基因表达分析表明，在简单培养系统中，BMP4 处理的细胞中生殖细胞特异性基因（VASA 和 Stra8）的表达比例显著更高。此外，在简单培养系统中的免疫细胞化学结果表明，与 BMP4 无处理组相比，VASA（原始生殖细胞（PGC）标志物）免疫染色细胞的平均百分比在 BMP4 处理组中显著增加。同时，CDH1，晚期精原生殖细胞标志物，在这两组之间没有显著差异。结果表明，BMP4 是从小鼠 ESC 中诱导 PGC 的有效诱导剂。然而，使用 MEFs 作为饲养层对 PGC 的诱导没有明显影响。
