Induction of dedifferentiated male mouse adipose stromal vascular fraction cells to primordial germ cell-like cells
Induction of dedifferentiated male mouse adipose stromal vascular fraction cells to primordial germ cell-like cells
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去分化雄性小鼠脂肪基质血管成分细胞诱导为原始生殖细胞样细胞
DOI：
10.1002/cbin.10305
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发表时间：
2014-09-01
影响因子：
3.9
通讯作者：
Guo, Xin
中科院分区：
文献类型：
--
作者：
Cui, Guanghui;Qi, Zhengyu;Guo, Xin
脂肪基质血管成分（SVF）含有丰富的间充质干细胞群，其自我更新和分化能力有限。通过重编程因子（c-Myc、Oct 4、Sox 2和Klf 4）使雄性小鼠脂肪SVF细胞去分化以形成胚胎干细胞样细胞（ESCLC），这提升了其有限的分化潜能。通过在分别提供有激活素A和BMP-4的培养基中培养，诱导ESCLC向上胚层样细胞（EpiLC）和原始生殖细胞样细胞（PGCLC）分化。体外培养的ESCLCs具有胚胎干细胞的特性，并能自动形成胚胎体。ESCLCs在EpiLC诱导培养基中培养2-3 d后，形成与其原始水滴样集落不同的扁平上皮结构，多能性相关基因表达降低。将在EpiLC诱导培养基中培养2天的细胞分离，并在PGCLC诱导培养基中培养4-6天，它们再次形成典型的水滴状集落。此外，多能性相关基因和原始生殖细胞（PGC）特异性相关基因的表达增加。在ESCLC向EpiLC和PGCLC的进展过程中，组蛋白甲基化酶H3 K9 me 2和H3 K27 me 3的水平不断变化，这与PGC特化中所见的相似。衍生的PGCLC表达SSEA-1、Blimp-1和Stella。此外，Igf 2 r和Snrpn的甲基化被保留，但H19和Kcnq 1 ot 1甲基化水平与非PGCLC相比略有降低，这表明衍生的PGCLC可能启动了印记擦除过程。
The adipose stromal vascular fraction (SVF) contains abundant mesenchymal stem cell populations that have a limited ability to self-renew and differentiate. Male mouse adipose SVF cells were dedifferentiated by reprogramming factors (c-Myc, Oct4, Sox2, and Klf4) to form embryonic stem cell-like cells (ESCLCs), which upgraded their limited differentiation potential. The ESCLCs were induced to differentiate toward epiblast-like cells (EpiLCs) and primordial germ cell-like cells (PGCLCs) by culturing in media supplied with activin A and BMP-4, respectively. The derived ESCLCs possess embryonic stem cell features and can automatically form embryonic bodies. After culture in EpiLC induction medium for 2-3 days, ESCLCs formed flattened epithelial structures that were different from their original water drop-like colonies, and the expression of pluripotency-related genes decreased. When the cells that had been cultured in EpiLC induction medium for 2 days were isolated and cultured in PGCLC induction medium for 4-6 days, they formed typical water drop-like colonies again. Moreover, expression of the pluripotency-related genes and the primordial germ cell (PGC) specification-related genes increased. During progression from ESCLCs toward EpiLCs and PGCLCs, the levels of histone methylases H3K9me2 and H3K27me3 kept changing, which resembled those seen in PGC specification. The derived PGCLCs expressed SSEA-1, Blimp-1, and Stella. Furthermore, methylation of Igf2r and Snrpn was retained, but H19 and Kcnq1ot1 methylation levels were slightly reduced compared to non-PGCLCs, suggesting that the derived PGCLCs may have initiated the process of imprint erasure.