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首页> 外文期刊>Tissue engineering, Part C. Methods >Agitation can induce differentiation of human pluripotent stem cells in microcarrier cultures.
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Agitation can induce differentiation of human pluripotent stem cells in microcarrier cultures.

机译：搅拌可在微载体培养物中诱导人多能干细胞分化。

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One of the factors that can impact human embryonic stem cell expansion in stirred microcarrier culture reactors is mechanical stress caused by agitation. Therefore, we have investigated the effects of agitation on human embryonic stem cell growth and expression of pluripotent markers. Agitation of HES-2 cell line in microcarrier cultures in stirred spinner and agitated six-well plates did not affect expression of pluripotent markers, cell viability, and cell doubling times even after seven passages. However, HES-3 cell line was found to be shear sensitive, showing downregulation of three pluripotent markers Oct-4, mAb 84, and Tra-1-60, and lower cell densities in agitated as compared with static cultures, even after one passage. Cell viability was unaffected. The HES-3-agitated cultures showed increased expression of genes and proteins of the three germ layers. We were unable to prevent loss of pluripotent markers or restore doubling times in agitated HES-3 microcarrier cultures by addition of five different known cell protective polymers. In addition, the human induced pluripotent cell line IMR90 was also shown to differentiate in agitated conditions. These results indicate that the effect of agitation on cell growth and differentiation is cell line specific. We assume that the changes in the growth and differentiation of the agitation-sensitive (HES-3) cell line do not result from the effect of shear stress directly on cell viability, but rather by signaling effects that influence the cells to differentiate resulting in slower growth.
机译：在搅拌的微载体培养反应器中影响人胚胎干细胞扩增的因素之一是搅拌引起的机械应力。因此，我们研究了搅拌对人类胚胎干细胞生长和多能标记表达的影响。即使在经过七次传代后，在搅拌器和搅拌的六孔板中的微载体培养中搅动HES-2细胞系也不会影响多能标记物的表达，细胞活力和细胞倍增时间。但是，发现HES-3细胞系具有剪切敏感性，即使在一次传代后，与静态培养相比，三种多能性标志物Oct-4，mAb 84和Tra-1-60的表达仍下降，并且搅拌下的细胞密度较低。 。细胞活力不受影响。 HES-3搅拌培养物显示了三个胚层的基因和蛋白质表达增加。我们无法通过添加五种不同的已知细胞保护性聚合物来防止多能标记的丢失或无法在搅拌的HES-3微载体培养物中恢复倍增时间。另外，人诱导的多能细胞系IMR90也显示在激动条件下分化。这些结果表明，搅拌对细胞生长和分化的影响是细胞系特异性的。我们假设搅动敏感（HES-3）细胞系的生长和分化的变化不是直接由剪切应力对细胞生存力的影响引起的，而是由影响细胞分化的信号传导作用导致的，导致细胞变慢增长。

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