Isolation, Culture, and Identification of Amniotic Fluid-Derived Mesenchymal Stem Cells
Isolation, Culture, and Identification of Amniotic Fluid-Derived Mesenchymal Stem Cells
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羊水间充质干细胞的分离、培养和鉴定
DOI：
10.1007/s12013-013-9558-z
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发表时间：
2013-11-01
影响因子：
2.6
通讯作者：
Ruiz, George
中科院分区：
文献类型：
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作者：
Fei, Xuetao;Jiang, Shan;Ruiz, George
羊水间充质干细胞（AF-MSC）是一种新发现的具有良好分化能力且易于获得的优良种子细胞。然而，重要的是开发一种有效分离和培养AF-MSC的方法。取兔羊水，贴壁法培养AF-MSC。采用流式细胞术、免疫印迹和免疫荧光法分析培养的AF-MSC的表型特征。成功地从羊水中分离培养了羊水间充质干细胞。流式细胞术分析表明，这些细胞表达CD 29和CD 44，而它们不表达CD 34。Western blot和免疫荧光分析证实了转录因子Oct-4的表达。使用粘附方法，我们开发了一个成功的，可重复的协议，从羊水中分离AF-MSC。表型分析结果显示，本研究分离的AF-MSC为多能细胞。
Amniotic fluid-derived mesenchymal stem cells (AF-MSC) are newly described, excellent seed cells that have good differentiation capability and are convenient to obtain. However, it is important to develop a method to isolate and culture AF-MSC efficiently. Amniotic fluid samples were obtained from rabbits and the adherence method was used for AF-MSC culture. Flow cytometry, western blot, and immunofluorescence studies were used to analyze the phenotypic characteristics of the cultured AF-MSC. Amniotic fluid-derived mesenchymal stem cells were successfully isolated and cultured from amniotic fluid. Flow cytometric analysis demonstrated that these cells expressed CD29 and CD44, while they did not express CD34. The expression of transcription factor Oct-4 was confirmed by western blot and immunofluorescence analysis. Using the adherence method, we developed a successful, reproducible protocol for the isolation of AF-MSC from amniotic fluid. The results of our phenotypic analysis revealed that the AF-MSC isolated in the present study were multipotent cells.