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首页> 美国卫生研究院文献>PLoS Clinical Trials >The Good into the Pot the Bad into the Crop!—A New Technology to Free Stem Cells from Feeder Cells
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The Good into the Pot the Bad into the Crop!—A New Technology to Free Stem Cells from Feeder Cells

机译：把好东西放进锅里把坏东西放进庄稼！ —一种从饲养细胞中释放干细胞的新技术

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A variety of embryonic and adult stem cell lines require an intial co-culturing with feeder cells for non-differentiated growth, self renewal and maintenance of pluripotency. However for many downstream ES cell applications the feeder cells have to be considered contaminations that might interfere not just with the analysis of experimental data but also with clinical application and tissue engineering approaches. Here we introduce a novel technique that allows for the selection of pure feeder-freed stem cells, following stem cell proliferation on feeder cell layers. Complete and reproducible separation of feeder and embryonic stem cells was accomplished by adaptation of an automated cell selection system that resulted in the aspiration of distinct cell colonies or fraction of colonies according to predefined physical parameters. Analyzing neuronal differentiation we demonstrated feeder-freed stem cells to exhibit differentiation potentials comparable to embryonic stem cells differentiated under standard conditions. However, embryoid body growth as well as differentiation of stem cells into cardiomyocytes was significantly enhanced in feeder-freed cells, indicating a feeder cell dependent modulation of lineage differentiation during early embryoid body development. These findings underline the necessity to separate stem and feeder cells before the initiation of in vitro differentiation. The complete separation of stem and feeder cells by this new technology results in pure stem cell populations for translational approaches. Furthermore, a more detailed analysis of the effect of feeder cells on stem cell differentiation is now possible, that might facilitate the identification and development of new optimized human or genetically modified feeder cell lines.
机译：多种胚胎干细胞和成年干细胞系需要与饲养细胞进行初始培养，以实现未分化生长，自我更新和维持多能性。但是，对于许多下游ES细胞应用，必须将饲养细胞视为污染物，这些污染物不仅可能干扰实验数据的分析，而且可能干扰临床应用和组织工程方法。在这里，我们介绍一种新技术，该技术允许在饲养细胞层上干细胞增殖后，选择无饲养细胞的纯干细胞。饲养细胞和胚胎干细胞的完全和可再现的分离是通过适应自动细胞选择系统完成的，该系统导致根据预定的物理参数吸引不同的细胞集落或部分集落。分析神经元分化，我们证明了无饲养干细胞表现出与在标准条件下分化的胚胎干细胞相当的分化潜能。然而，无饲养层的细胞中胚状体的生长以及干细胞向心肌细胞的分化显着增强，这表明在早期胚状体发育过程中，依赖于饲养层的细胞对谱系分化进行了调节。这些发现强调了在体外分化开始之前分离干细胞和饲养细胞的必要性。通过这项新技术将干细胞和饲养细胞完全分离，可以得到纯干细胞群体进行翻译。此外，现在有可能对饲养细胞对干细胞分化的影响进行更详细的分析，这可能有助于鉴定和开发新的优化的人类或基因修饰的饲养细胞系。

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