Biochemistry Department, Shiraz University of Medical Sciences, Medical School, Shiraz, Iran.
Mol Biol Rep. 2012 Jul;39(7):7339-46. doi: 10.1007/s11033-012-1565-4.
Generation of patient specific stem cells is among the ultimate goals in regenerative medicine. Such a cell needs to be functional when it transplants. Interaction between the matrix proteins and integrin adjust many cells' function such as adhesion, migration, cell cycle and self renewal in stem cells. In this study, NIH3T3 cells were dedifferentiated by mouse Embryonic Stem Cell (mESC) extract. The expression of pluripotency markers as well as a2, a5 and a6 integrin subunits were determined. NIH3T3 cells treated with mESC extract showed noticeable changes in cell morphology as early as day 2 post-treatment forming colonies similar to typical mESC morphology by day 8, after three passages. Alkaline phosphatase (ALP) assay and immunocytochemistry staining were performed for the induced reprogrammed cells. The results indicated that these colonies showed the ALP activity and they express Sox2 and Nanog. RT-PCR revealed that the colonies also express Oct3/4. NIH3T3 cells, ESC and reprogrammed cells expressed a2 integrin. a5 integrin expression was greatest in reprogrammed cells followed by the expression of this integrin in NIH3T3 which in turn was more than in ESC. a6A integrin was expressed in NIH3T3 cells while a6B integrin was expressed in ESC and in very low quantity was expressed in reprogrammed cells. These data provide evidence for both the generation of ES like cells from differentiated somatic cells and the expression profile of integrins after de-differentiation by mESC extract.
生成患者特异性干细胞是再生医学的最终目标之一。这种细胞在移植时需要具有功能。基质蛋白与整合素之间的相互作用调节许多细胞的功能，如干细胞的黏附、迁移、细胞周期和自我更新。在这项研究中，NIH3T3 细胞通过鼠胚胎干细胞（mESC）提取物去分化。确定多能性标记物以及整合素 a2、a5 和 a6 亚基的表达。NIH3T3 细胞用 mESC 提取物处理后，在处理后第 2 天，细胞形态发生明显变化，形成类似于典型 mESC 形态的集落，到第 8 天，经过 3 次传代后。对诱导重编程细胞进行碱性磷酸酶（ALP）测定和免疫细胞化学染色。结果表明，这些集落表现出 ALP 活性，并且它们表达 Sox2 和 Nanog。RT-PCR 显示这些集落还表达 Oct3/4。NIH3T3 细胞、ESC 和重编程细胞表达 a2 整合素。a5 整合素在重编程细胞中的表达最强，其次是 NIH3T3 细胞中的表达，而 ESC 中的表达则更高。a6A 整合素在 NIH3T3 细胞中表达，而 a6B 整合素在 ESC 中表达，在重编程细胞中表达量很低。这些数据为从分化的体细胞生成类似 ES 的细胞以及 mESC 提取物去分化后整合素的表达谱提供了证据。
