Development of a perfusion fed bioreactor for embryonic stem cell-derived cardiomyocyte generation: Oxygen-mediated enhancement of cardiomyocyte output
Development of a perfusion fed bioreactor for embryonic stem cell-derived cardiomyocyte generation: Oxygen-mediated enhancement of cardiomyocyte output
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DOI：
10.1002/bit.20445
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发表时间：
2005-05-20
影响因子：
3.8
通讯作者：
Zandstra, PW
中科院分区：
文献类型：
--
作者：
Bauwens, C;Yin, T;Zandstra, PW
细胞移植是一种很有前途的新方法，以取代疤痕，无功能的心肌在患病的心脏。然而，目前，产生开发和测试动物模型所需的供体心肌细胞的数量是一个主要的限制。胚胎干细胞（ES）可能是治疗应用的一个有前途的来源，可能提供足够数量的功能相关的细胞移植到各种器官。我们开发了一种用于ES细胞衍生的心肌细胞生产的单步生物过程，该过程能够实现培养基灌注和溶解氧的直接监测和控制。生物过程的实施需要结合方法来防止ES细胞聚集（水凝胶包封）和从异质细胞群中纯化心肌细胞（遗传选择），在受控的生物反应器环境中进行培养基灌注。我们利用这一生物过程来研究氧对心肌细胞生成的影响。平行容器（250 mL培养体积）在常氧（20%氧张力）或低氧（4%氧张力）条件下运行。经过14天的分化（包括5天的选择），在缺氧血管中获得的每输入ES细胞的心肌细胞产量为3.77 +/- 0.13，高于先前报道的。我们已经开发了一种生物过程，提高了ES细胞衍生的心肌细胞生产的效率，并允许对ES细胞衍生的心肌发生的生物过程参数的调查。使用这个系统，我们已经证明，中等氧张力是一个培养参数，可以操纵，以提高心肌细胞产量。(c)2005 Wiley Periodicals，Inc.
Cell transplantation is emerging as a promising new approach to replace scarred, nonfunctional myocardium in a diseased heart. At present, however, generating the numbers of donor cardiomyocytes required to develop and test animal models is a major limitation. Embryonic stem (ES) cells may be a promising source for therapeutic applications, potentially providing sufficient numbers of functionally relevant cells for transplantation into a variety of organs. We developed a single-step bioprocess for ES cell-derived cardiomyocyte production that enables both medium perfusion and direct monitoring and control of dissolved oxygen. Implementation of the bioprocess required combining methods to prevent ES cell aggregation (hydrogel encapsulation) and to purify for cardiomyocytes from the heterogeneous cell populations (genetic selection), with medium perfusion in a controlled bioreactor environment. We used this bioprocess to investigate the effects of oxygen on cardiomyocyte generation. Parallel vessels (250 mL culture volume) were run under normoxic (20% oxygen tension) or hypoxic (4% oxygen tension) conditions. After 14 days of differentiation (including 5 days of selection), the cardiomyocyte yield per input ES cell achieved in hypoxic vessels was 3.77 +/- 0.13, higher than has previously been reported. We have developed a bioprocess that improves the efficiency of ES cell-derived cardiomyocyte production, and allows the investigation of bioprocess parameters on ES cell-derived cardiomyogenesis. Using this system we have demonstrated that medium oxygen tension is a culture parameter that can be manipulated to improve cardiomyocyte yield. (c) 2005 Wiley Periodicals, Inc.