Li N, Pan S, Zhu H, Mu H, Liu W, Hua J
College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology, Key Lab for Animal Biotechnology of Agriculture Ministry of China, Northwest A&F University, Yangling, Shaanxi, 712100, China.
Cell Prolif. 2014 Aug;47(4):299-309. doi: 10.1111/cpr.12115. Epub 2014 Jun 13.
Recent studies have demonstrated that primordial germ cells (PGC) can be differentiated from human umbilical cord mesenchymal stem cells (hUC-MSCs), and embryonic stem cells (ESCs) in vitro. Nevertheless, efficiencies were low and unstable. Here, whether hUC-MSCs can be induced to differentiate into germ-like cells with the aid of bone morphogenetic protein (BMP4) was investigated.
Human umbilical cord mesenchymal stem cells were freshly isolated and cultured with BMP4. SSEA-1(+/-) cells were purified using magnetic-activated cell sorting (MACS) from the hUC-MSCs, and further induced with BMP4. Quantitative real-time PCR (qRT-PCR) and immunofluorescence analysis were used to determine PGC and germ-like cell-specific markers.
Human umbilical cord mesenchymal stem cells differentiated into SSEA-1(+) spherical PGC-like cells efficiently with 12.5 ng/ml BMP4. qRT-PCR and immunofluorescence analysis demonstrated that SSEA-1(+) cells expressed higher levels of PGC-specific markers than SSEA-1(-) cells. Furthermore, SSEA-1(+) cells were induced with BMP4 to differentiate into STRA8, SCP3, DMRT1 and PLZF-positive male germ-like cells, and some sperm-like cells were obtained by 7-14 days after induction.
These results suggest that SSEA-1(+) hUC-MSCs can differentiate into male germ-like cells in the presence of BMP4. This study provides an efficient protocol to study germ-cell development using hUC-MSCs.
近期研究表明，原始生殖细胞（PGC）可在体外由人脐带间充质干细胞（hUC-MSCs）和胚胎干细胞（ESCs）分化而来。然而，分化效率较低且不稳定。在此，研究了hUC-MSCs在骨形态发生蛋白（BMP4）的辅助下是否可被诱导分化为类生殖细胞。
新鲜分离人脐带间充质干细胞并用BMP4进行培养。使用磁珠分选法（MACS）从hUC-MSCs中纯化出阶段特异性胚胎抗原-1（SSEA-1）（+/-）细胞，并用BMP4进一步诱导。采用定量实时聚合酶链反应（qRT-PCR）和免疫荧光分析来确定PGC和类生殖细胞特异性标志物。
人脐带间充质干细胞在12.5 ng/ml BMP4作用下高效分化为SSEA-1（+）球形类PGC细胞。qRT-PCR和免疫荧光分析表明，SSEA-1（+）细胞比SSEA-1（-）细胞表达更高水平的PGC特异性标志物。此外，用BMP4诱导SSEA-1（+）细胞分化为STRA8、SCP3、DMRT1和PLZF阳性的雄性类生殖细胞，诱导7 - 14天后获得了一些精子样细胞。
这些结果表明，在BMP4存在的情况下，SSEA-1（+）hUC-MSCs可分化为雄性类生殖细胞。本研究提供了一种利用hUC-MSCs研究生殖细胞发育的有效方案。
