In vitro differentiation of male mouse embryonic stem cells into both presumptive sperm cells and oocytes
In vitro differentiation of male mouse embryonic stem cells into both presumptive sperm cells and oocytes
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DOI：
10.1089/clo.2007.0031
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发表时间：
2007-12-01
期刊：
影响因子：
--
通讯作者：
Kerkis, Irina
中科院分区：
文献类型：
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作者：
Kerkis, Alexandre;Fonseca, Simone A. S.;Kerkis, Irina
在雄性小鼠胚胎干细胞（ES）分化为生殖细胞（GC）方面的开创性工作表明，在单独的实验中，利用经过基因操作或预先选择的ES细胞可产生雄性或雌性配子。为了在不进行任何基因操作或预先选择的情况下从雄性小鼠ES细胞产生两种类型的配子，我们通过视黄酸（RA）在非贴壁拟胚体（EB）内诱导分化。基于诸如Oct - 4、Mvh、Stella、Dazl、Piwil 2、Pdrd 1、Rex 14、Rnf 17、Bmp8b、顶体素（Acrosin）、Stra - 8、Haprin、LH - R、Gdp、Zp3、Zp2、Sycp1和Sycp3等早期和晚期生殖细胞特异性基因的表达，似乎配子样细胞以正确的方式形成。对形态良好的生殖细胞和假定配子进行的免疫荧光分析显示，SSEA1、Oct - 4、EMA - 1、FE - J1、Dazl、Fragilis、Mvh、顶体素和乙酰化α - 微管蛋白呈阳性标记。常规细胞遗传学和荧光原位杂交（FISH）分析表明，ES来源的生殖细胞存在染色体减少。我们的数据表明，具有XY染色体的ES细胞在相同的实验条件下能够产生两种类型的假定配子，并且这种产生取决于它们在拟胚体环境中的位置和时间信息。
Pioneer work in male mouse embryonic stem (ES) cells differentiation into germ cells (GC) showed generations of male or female gametes in separate experiments, using genetically manipulated or preselected ES cells. In an attempt to produce both types of gametes from male mouse ES cells without any genetic manipulation or preselection, we induce the differentiation by retinoic acid (RA) within nonadherent embryoid bodies (EB). It seems that gamete-like cell formation occurs in the correct manner based on the expression of early and late GC-specific genes such as Oct-4, Mvh, Stella, Dazl, Piwil 2, Pdrd 1, Rex 14, Rnf 17, Bmp8b, Acrosin, Stra-8, Haprin, LH-R, Gdp, Zp3, Zp2, Sycp1, and Sycp3. Immunofluorescence analysis of morphologically well-formed GC and presumptive gametes showed positive labeling for SSEA1, Oct-4, EMA-1, FE-J1, Dazl, Fragilis, Mvh, Acrosin, and acetylated a-tubulin. Conventional cytogenetic and FISH analysis indicated a chromosome reduction in ES-derived GC. Our data suggest that ES cells with XY. chromosomes can produce under the same experimental conditions both types of presumptive gametes, and this production depends on their positional and temporal information within the EB context.