BMP4 promotes SSEA-1+ hUC-MSC differentiation into male germ-like cells in vitro
BMP4 promotes SSEA-1+ hUC-MSC differentiation into male germ-like cells in vitro
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DOI：
10.1111/cpr.12115
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发表时间：
2014-08-01
影响因子：
8.5
通讯作者：
Hua, J.
中科院分区：
文献类型：
--
作者：
Li, N.;Pan, S.;Hua, J.
目的：最近的研究表明，人脐带间充质干细胞（hUC-MSC）和胚胎干细胞（ESCs）可以在体外分化为原始生殖细胞（PGC）。然而，效率低下且不稳定。在这里，是否hUC-MSCs可以诱导分化为生殖样cells与骨形态发生蛋白（BMP 4）的援助investigated.Materials和方法：人脐带间充质干细胞新鲜分离和培养与BMP 4。使用磁激活细胞分选（MACS）从hUC-MSC纯化SSEA-1（+/-）细胞，并进一步用BMP 4诱导。结果：人脐带间充质干细胞在12.5 ng/ml BMP 4的诱导下，可高效分化为SSEA-1（+）球形PGC样细胞。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。结论：SSEA-1（+）hUC-MSCs经BMP 4诱导后可分化为STRA 8、SCP 3、DMRT 1和PLZF阳性的雄性生殖样细胞，诱导后7-14 d可获得部分精子样细胞。本研究提供了一个有效的方案，研究生殖细胞的发展，使用hUC-MSCs。
Objectives: 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.Materials and methods: 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.Results: 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.Conclusion: 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.