Bioreactor cultivation enhances the efficiency of human embryoid body (hEB) formation and differentiation
Bioreactor cultivation enhances the efficiency of human embryoid body (hEB) formation and differentiation
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DOI：
10.1002/bit.20045
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发表时间：
2004-06-05
影响因子：
3.8
通讯作者：
Itskovitz-Eldor, J
中科院分区：
文献类型：
--
作者：
Gerecht-Nir, S;Cohen, S;Itskovitz-Eldor, J
人胚胎干细胞（hESC）为细胞治疗和组织工程提供无限细胞供应的前景取决于其扩增和分化的可控生物过程的可用性。我们首次描述了在旋转生物反应器中形成分化的人胚状体（hEBs），以尝试控制其团聚。相对于hEB形成和分化的产率，分析了动态过程与培养皿中静态培养相比的功效。定量分析的hEB，DNA和蛋白质含量，和活细胞浓度，作为培养细胞和规模的措施，揭示了3倍的增强生成的hEB相比，静态培养。其他代谢指标，如葡萄糖消耗，乳酸产生，和pH值指出，有效的细胞扩增和分化的动态培养。旋转容器的类型对hEB的形成和团聚过程有显著影响。在慢转侧血管（STLV），hEB的大小较小，没有看到大的坏死中心，即使在1个月的培养。在高纵横比旋转容器（HARV）中，hEB聚集是大量的。来自三个胚层的代表性组织的外观以及原始神经元管组织、血管形成和特异性内分泌表明，在动态形成的hEB中，初始发育事件没有改变。总的来说，我们的研究定义了培养条件，其中获得对分化中的hESC的聚集的控制，从而使临床和工业应用的可规模化细胞生产成为可能。(C)2004 Wiley Periodicals，Inc.
The promise of human embryonic stem cells (hESCs) to provide an unlimited supply of cells for cell therapy and tissue engineering depends on the availability of a controllable bioprocess for their expansion and differentiation. We describe for the first time the formation of differentiating human embryoid bodies (hEBs) in rotating bioreactors to try and control their agglomeration. The efficacy of the dynamic process compared to static cultivation in Petri dishes was analyzed with respect to the yield of hEB formation and differentiation. Quantitative analyses of hEBs, DNA and protein contents, and viable cell concentration, as measures for culture cellularity and scale-up, revealed 3-fold enhancement in generation of hEBs compared to the static culture. Other metabolic indices such as glucose consumption, lactic acid production, and pH pointed to efficient cell expansion and differentiation in the dynamic cultures. The type of rotating vessel had a significant impact on the process of hEB formation and agglomeration. In the slow turning lateral vessel (STLV), hEBs were smaller in size and no large necrotic centers were seen, even after 1-month cultivation. In the high aspect rotating vessel (HARV), hEB agglomeration was massive. The appearance of representative tissues derived from the three germ layers as well as primitive neuronal tube organization, blood vessel formation, and specific-endocrine secretion indicated that the initial developmental events are not altered in the dynamically formed hEBs. Collectively, our study defines the culture conditions in which control over the aggregation of differentiating hESCs is obtained, thus enabling scaleable cell production for clinical and industrial applications. (C) 2004 Wiley Periodicals, Inc.