Primordial germ cell-like cells differentiated in vitro from skin-derived stem cells.
Primordial germ cell-like cells differentiated in vitro from skin-derived stem cells.
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DOI：
10.1371/journal.pone.0008263
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发表时间：
2009-12-14
期刊：
影响因子：
3.7
通讯作者：
Li J
中科院分区：
文献类型：
--
作者：
Linher K;Dyce P;Li J
我们先前已经证明，从胎猪皮肤中分离的干细胞具有在体外形成卵母细胞样细胞（OLC）的潜力。然而，原始生殖细胞（PGCs），这也必须指定在干细胞分化过程中产生这些假定的卵母细胞在更先进的阶段的文化，没有系统的特点。目前的研究测试了一个假设，即在OLC形成之前，来自皮肤干细胞的形态学上不同的细胞群对应于假定的PGC，其进一步分化为更成熟的配子。当在适当的微环境中诱导分化时，形态上不同的细胞亚群（其中一些是碱性磷酸酶（AP）阳性的）也表达Oct 4、Fragilis、Stella、Dazl和Vasa，它们是指示生殖细胞形成的标志物。一个已知的差异甲基化区域（DMR）内的H19基因座，这是去甲基化的卵母细胞后，建立的母亲的印记，是低甲基化的PGC样细胞相比，未分化的皮肤来源的干细胞，这表明假定的生殖细胞群体经历印记擦除。支持体外产生的PGC样细胞的生殖细胞身份的另外证据是，当用Dazl-GFP报告基因标记时，这些细胞进一步分化成GFP阳性OLC。从体干细胞产生生殖细胞前体的能力可能提供一种体外模型来研究有关早期生殖细胞形成的一些未解问题。
We have previously demonstrated that stem cells isolated from fetal porcine skin have the potential to form oocyte-like cells (OLCs) in vitro. However, primordial germ cells (PGCs), which must also be specified during the stem cell differentiation to give rise to these putative oocytes at more advanced stages of culture, were not systematically characterized. The current study tested the hypothesis that a morphologically distinct population of cells derived from skin stem cells prior to OLC formation corresponds to putative PGCs, which differentiate further into more mature gametes. When induced to differentiate in an appropriate microenvironment, a subpopulation of morphologically distinct cells, some of which are alkaline phosphatase (AP)-positive, also express Oct4, Fragilis, Stella, Dazl, and Vasa, which are markers indicative of germ cell formation. A known differentially methylated region (DMR) within the H19 gene locus, which is demethylated in oocytes after establishment of the maternal imprint, is hypomethylated in PGC-like cells compared to undifferentiated skin-derived stem cells, suggesting that the putative germ cell population undergoes imprint erasure. Additional evidence supporting the germ cell identity of in vitro-generated PGC-like cells is that, when labeled with a Dazl-GFP reporter, these cells further differentiate into GFP-positive OLCs. The ability to generate germ cell precursors from somatic stem cells may provide an in vitro model to study some of the unanswered questions surrounding early germ cell formation.
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影响因子：
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作者：
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发表时间：
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