Derivation and maintenance of murine trophoblast stem cells under defined conditions.
Derivation and maintenance of murine trophoblast stem cells under defined conditions.
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DOI：
10.1016/j.stemcr.2013.12.013
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发表时间：
2014-02-11
影响因子：
5.9
通讯作者：
Schorle, Hubert
中科院分区：
文献类型：
--
作者：
Kubaczka, Caroline;Senner, Claire;Arauzo-Bravo, Marcos J.;Sharma, Neha;Kuckenberg, Peter;Becker, Astrid;Zimmer, Andreas;Bruestle, Oliver;Peitz, Michael;Hemberger, Myriam;Schorle, Hubert
滋养层干细胞（TSCs）是胎盘前体细胞的体外等同物。将TSC培养在含有成纤维细胞生长因子4、肝素和胚胎成纤维细胞条件培养基的富血清培养基中。在这里，我们开发了一种简单的培养基，由10种化学成分确定的成分组成，用于在Matrigel或合成基质上培养TSCs，称为TX培养基。基因表达和DNA甲基化分析证明了在TX中培养的TSC的表达谱和表观基因组特征的忠实繁殖。此外，TX培养基支持TSC系的从头衍生。最后，在TX中培养的TSC在体外分化成滋养外胚层谱系的所有衍生物，在裸鼠中引起出血性病变，并嵌合胎盘，表明它们保留了TSC的所有特征。TX培养基配方不再需要胎牛血清和条件培养基，这有利于并简化了这种胚外谱系的培养。TSC可以在基质胶包被的表面上的确定的无血清TX培养基中生长。在TX培养中，维持干细胞标志物的表达和稳定的核型TX中的表达和甲基化谱与标准条件高度相似体外和体内TX培养基中的TSC保持完全分化潜能Schorle和同事重新研究了TSCs的分化潜能。研究了胚胎外滋养层干细胞生长的要求，并证明了十种化学成分可以取代胎牛血清和成纤维细胞条件培养基，过去不一致的主要来源。滋养层干细胞可以在该培养基中衍生、增殖和分化。这极大地促进了TS研究，因为它为这种类型的实验提供了一个标准化的平台。
Trophoblast stem cells (TSCs) are in vitro equivalents to the precursor cells of the placenta. TSCs are cultured in serum-rich medium with fibroblast growth factor 4, heparin, and embryonic-fibroblast-conditioned medium. Here, we developed a simple medium consisting of ten chemically defined ingredients for culture of TSCs on Matrigel or synthetic substrates, named TX medium. Gene expression and DNA methylation profiling demonstrated the faithful propagation of expression profiles and epigenomic characteristics of TSCs cultured in TX. Further, TX medium supported the de novo derivation of TSC lines. Finally, TSCs cultured in TX differentiate into all derivatives of the trophectodermal lineage in vitro, give rise to hemorrhagic lesions in nude mice, and chimerize the placenta, indicating that they retained all hallmarks of TSCs. TX media formulation no longer requires fetal bovine serum and conditioned medium, which facilitates and standardizes the culture of this extraembryonic lineage. TSCs can be grown in defined serum-free TX media on Matrigel-coated surfaces In TX culture, expression of stem cell markers and a stable karyotype are maintained Expression and methylation profiles in TX are highly similar to standard conditions TSCs retain full differentiation potential in TX media in vitro and in vivo Schorle and colleagues re-examined the requirements for the growth of extraembryonic trophoblast stem cells and demonstrate that ten chemical defined ingredients can replace fetal bovine serum and fibroblast-conditioned media, major sources of inconsistencies in the past. Trophoblast stem cells can be derived, propagated, and differentiated in this medium. This greatly facilitates TS research because it provides a standardized platform for this type of experiments.
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影响因子：
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作者：
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影响因子：
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作者：
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通讯作者：
Smith, Austin
影响因子：
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作者：
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通讯作者：
Winterhager, E