Lowering Oxygen Tension Enhances the Differentiation of Mouse Embryonic Stem Cells into Neuronal Cells
Lowering Oxygen Tension Enhances the Differentiation of Mouse Embryonic Stem Cells into Neuronal Cells
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DOI：
10.1002/btpr.248
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发表时间：
2009-09-01
影响因子：
2.9
通讯作者：
Veraitch, Farlan S.
中科院分区：
文献类型：
--
作者：
Mondragon-Teran, Paul;Lye, Gary J.;Veraitch, Farlan S.
胚胎干细胞（ESC）能够在体外无限增殖，同时保留其分化为每个成体谱系细胞的能力。高效、高产的方法，将ESC分化为特定的谱系，将为开发具有成本效益的药物筛选和细胞治疗产品奠定基础。本研究的目的是调查目前用于ESC神经元分化的实验室氧张力是否是次优的，从而导致低效的过程产量。在氧气控制的室中使用化学成分确定的培养基进行用于小鼠ESC（mESC）的神经元分化的贴壁单层方案，培养8天。当暴露于更适合于体内神经元发育的氧张力（2%O（2））时，分化过程中活细胞的产量增加了34倍。低氧张力抑制细胞死亡过程中的早期阶段（48至96小时）和接近尾声（120至192小时）。通过流式细胞术测定表达神经元标记物的细胞百分比，显示在2%O（2）下β III微管蛋白的小幅上升和MAP 2群体的三倍增加。表达神经元标志物的活细胞的产量的总增加显示为β III微管蛋白的55倍和MAP 2的114倍。总之，这项研究表明，低氧张力可用于提高从胚胎干细胞衍生的神经元细胞的产量，并对开发高效，具有成本效益的生产工艺具有影响。(C)2009年美国化学工程师学会生物技术。程序：25：1480-1488，2009
Embryonic stem cells (ESC) are capable of proliferating indefinitely in vitro whilst retaining their ability to differentiate into cells of every adult lineage. Efficient, high yield processes, which direct differentiation of ESC to specific lineages, will underpin the development of cost-effctive drug screening and cell therapy products. The aim of this study was to investigate whether laboratory oxygen tension currently used for the neuronal differentiation of ESC was suboptimal resulting in inefficient process yields. An adherent monolayer protocol for the neuronal differentiation of mouse ESC (mESC) was performed in oxygen controlled chambers using a chemically defined media over an 8 day period of culture. When exposed to oxygen tensions more appropriate to in vivo neuronal development (2% O(2)), there was a 34-fold increase in the yield of viable cells from the differentiation process. Low oxygen tension inhibited cell death during an early phase (48 to 96 h) and toward the end (120 to 192 h) of the process. The percentage of cells expressing neuronal markers was determined by flow cytometry, revealing a small rise in the beta III tubulin and a threefold increase in the MAP2 populations at 2% O(2). The total increase in the yield of viable cells expressing neuronal markers was shown to be 55-fold for beta III tubulin and 114-fold for MAP2. In conclusion, this study revealed that low oxygen tension can be used to enhance the yield of neuronal cells derived from ESCs and has implications for the development of efficient, cost-effective production processes. (C) 2009 American Institute of Chemical Engineers Biotechnol. Prog., 25: 1480-1488, 2009