【摘要】：目的比較OCT4異構體(OCT4A、OCT4B、OCT4B1)及其調控因子在人胚胎干細胞(hESC)和人間充質干細胞(hMSC)中的表達。方法利用RT-PCR、免疫熒光染色、流式細胞分析及體內/外分化實驗,鑒定hESC及hMSC的生物學特性;應用real-time PCR、Western blot和流式細胞分析比較OCT4異構體及其轉錄因子NANOG,SOX2和mRNA結合蛋白LIN28在hESC及hMSC中的表達水平。結果 OCT4異構體mRNA在hESC和hMSC中均有表達,在hESC中的表達顯著高于在hMSC中,并以OCT4A的差別最為顯著(P0.01);在蛋白水平,hESC表達OCT4A和OCT4B-256aa,hMSC不表達OCT4異構體蛋白。hESC高表達OCT4的調控因子NANOG、SOX2和LIN28;hMSC低表達SOX2,不表達NANOG和LIN28。結論 NANOG、SOX2和LIN28調控OCT4的表達,OCT4異構體在hESC和hMSC中的表達差異提示其可能是不同發(fā)育階段干細胞自我更新和分化潛能等方面差別的主要因素之一。
[Abstract]:Objective to compare the expression of OCT4 isomer (OCT4AnOCT4B1) and its regulatory factors in human embryonic stem cell (hESC) and human mesenchymal stem cell (hMSC). Methods the biological characteristics of hESC and hMSC were identified by RT-PCR, immunofluorescence staining, flow cytometry and in vitro / in vivo differentiation assay. The expression levels of OCT4 isomer and its transcription factor NANOGN SOX2 and mRNA binding protein LIN28 in hESC and hMSC were compared by real-time PCR Western blot and flow cytometry. Results OCT4 isomer mRNA was expressed in both hESC and hMSC, and the expression in hESC was significantly higher than that in hMSC. The expression of OCT4A and OCT4B-256aahMSC did not express OCT4 isomer protein HESC and the regulatory factors of high expression of OCT4, NANOGG SSOX2 and LIN28hMSC, but not NANOG and LIN28hMSC. Conclusion the difference in the expression of OCT4 isomers in hESC and hMSC suggests that NANOGN SOX2 and LIN28 may be one of the main factors of the difference in self-renewal and differentiation potential of stem cells in different developmental stages.
【作者單位】： 中國醫(yī)學科學院整形外科醫(yī)院研究中心;
【基金】：國家自然科學基金(31201102,30871433,31071305)
【分類號】：R329

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