Mix11 and Oct4 proteins are transiently co-expressed in differentiating mouse and human embryonic stem cells
Mix11 and Oct4 proteins are transiently co-expressed in differentiating mouse and human embryonic stem cells
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DOI：
10.1089/scd.2005.14.656
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发表时间：
2005-12-01
影响因子：
4
通讯作者：
Elefanty, AG
中科院分区：
文献类型：
--
作者：
Mossman, AK;Sourris, K;Elefanty, AG
胚胎干细胞(ESCs)具有形成体内所有组织的能力，因此，沿着特定谱系定向分化的ESCs是一种产生治疗有用细胞类型的手段。据推测，在体外分化过程中，ES细胞会依次经历与胚胎细胞相似的发育阶段。识别这些阶段的试剂的可用性将有助于监测和优化胚胎干细胞分化。一个关键的阶段是早期胚胎中内胚层和中胚层前体的发育，标志着转录因子Mixl1和干细胞基因Oct4的瞬时表达。为了在胚胎干细胞分化过程中识别相应的细胞，我们产生了针对Mixl1蛋白的单抗，该抗体可以很好地检测到老鼠和人类的蛋白质。细胞内流式细胞术显示，大约90%的分化小鼠胚胎干细胞瞬时共表达Oct4和Mixl1蛋白，部分分化的人类ES细胞也共表达MIXL1和OCT4蛋白。这些实验首次通过蛋白质表达证明，人和小鼠的ESCs在体外分化过程中都经历了与哺乳动物早期发育相对应的发育阶段。此外，这些研究证实，抗Mixl1抗体是监测ESC分化的有价值的试剂，并将有助于有效地产生临床相关的细胞类型。
Embryonic stem cells (ESCs) have the capacity to form all the tissues in the body and hence, directed differentiation of ESCs along specific lineages represents a means to generate therapeutically useful cell types. It has been postulated that, during in vitro differentiation, ES cells sequentially pass through similar developmental stages as cells in the embryo. The availability of reagents that identify these stages would facilitate the monitoring and optimization of ESC differentiation. One key stage, the development of endodermal and mesodermal precursors in the early embryo, is marked by the transient expression of the transcription factor, Mixl1 and the stem cell gene, Oct4. In order to identify corresponding cells during ESC differentiation, we generated monoclonal antibodies to the Mixl1 protein that robustly detected both mouse and human proteins. Intracellular flow cytometry was used to show that approximately 90% of differentiating mouse ESCs transiently co-expressed Oct4 and Mixl1 proteins and that a subset of differentiating human ES cells also co-expressed MIXL1 and OCT4 proteins. These experiments have demonstrated for the first time by protein expression that both human and mouse ESCs passed through developmental stages during in vitro differentiation that corresponded to those observed in early mammalian development. Furthermore, these studies confirmed that anti-Mixl1 antibodies are a valuable reagent for monitoring ESC differentiation and will facilitate the efficient generation of clinically relevant cell types.