Involvement of tyrosine kinase signaling in maintaining murine embryonic stem cell functionality
Involvement of tyrosine kinase signaling in maintaining murine embryonic stem cell functionality
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DOI：
10.1016/j.exphem.2007.04.010
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发表时间：
2007-08-01
影响因子：
2.6
通讯作者：
Helgason, Cheryl D.
中科院分区：
文献类型：
--
作者：
Lu, Min;Glover, Clive H.;Helgason, Cheryl D.
Objective.我们以前证明，c-kit表达减少小鼠胚胎干细胞（ESC）诱导的白血病抑制因子去除分化。在这项研究中，我们解决了c-kit是未分化的小鼠ESC的标志物的可能性，而且，它在维持这些细胞的未分化状态中发挥作用。通过流式细胞术和定量逆转录聚合酶链反应分析不同分化条件下的c-kit表达。然后根据c-kit表达对ESC进行分选，并使用胚状体和集落形成细胞测定研究其功能。以伊马替尼（Gleevec）和ACK 2阻断c-kit活性，以干细胞因子刺激c-kit活性。在不同分化条件下，两种鼠ESC系中c-kit表达降低。基于c-kit表达的ESC群体分选揭示了分选群体的功能能力和基因表达谱的显著差异。抑制研究揭示了酪氨酸激酶活性在维持ESC活力和分化能力中的重要作用，至少部分通过防止凋亡和增强细胞周期进程。然而，单独激活c-kit并不足以维持未分化的ESC。结果表明，c-kit可能是一个有用的标志物，用于监测ESC功能。此外，酪氨酸激酶信号在维持未分化ESC中起重要作用。这项工作提供了有价值的见解复杂的信号通路，协同维持未分化状态的小鼠ESC。(c)2007 ISEH -血液学和干细胞学会。爱思唯尔公司出版
Objective. We previously demonstrated that c-kit expression decreases during murine embryonic stem cell (ESC) differentiation induced by leukemia inhibitory factor removal. In this study, we addressed the possibility that c-kit is a marker of undifferentiated murine ESC and, moreover, that it plays a role in maintaining the undifferentiated state of these cells.Materials and Methods. c-kit expression was analyzed under various differentiation conditions by flow cytometry and quantitative reverse transcription polymerase chain reaction. ESC were then sorted on the basis of c-kit expression and functionality was investigated using embryoid body and colony-forming cell assays. Imatinib (Gleevec) and ACK2 were used to block, and stem cell factor was used to stimulate, c-kit activity.Results. c-kit expression decreased in two murine ESC lines under various differentiation conditions. Sorting of ESC populations on the basis of c-kit expression revealed significant differences in the functional capacities and gene expression profiles of the sorted populations. The inhibition studies revealed an important role for tyrosine kinase activity in maintaining ESC viability and differentiation capacity, at least in part by preventing apoptosis and enhancing cell cycle progression. However, activation of c-kit alone is not sufficient for maintaining undifferentiated ESC.Conclusion. The results suggest that c-kit may represent a useful marker for monitoring ESC functionality. Moreover, tyrosine kinase signaling plays an important role in maintaining undifferentiated ESC. This work provides valuable insights into the complex signaling pathways that synergize to maintain the undifferentiated state of murine ESC. (c) 2007 ISEH - Society for Hematology and Stem Cells. Published by Elsevier Inc.