Combining hypoxia and bioreactor hydrodynamics boosts induced pluripotent stem cell differentiation towards cardiomyocytes.
Combining hypoxia and bioreactor hydrodynamics boosts induced pluripotent stem cell differentiation towards cardiomyocytes.
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DOI：
10.1007/s12015-014-9533-0
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发表时间：
2014-12
影响因子：
4.8
通讯作者：
Alves, Paula M.
中科院分区：
文献类型：
--
作者：
Correia, Claudia;Serra, Margarida;Espinha, Nuno;Sousa, Marcos;Brito, Catarina;Burkert, Karsten;Zheng, Yunjie;Hescheler, Juergen;Carrondo, Manuel J. T.;Saric, Tomo;Alves, Paula M.
关键词：
从诱导多能干细胞(IPSCs)来源的心肌细胞(CMS)在针对患者的疾病模型、药物筛选和细胞治疗方面具有很大的前景。然而，现有的IPSC的CM分化方案除了高度依赖昂贵的生长因子的应用外，还显示出低的重复性和可扩展性。这项工作的目的是通过设计确保低氧和机械环境的生物反应器协议，开发一种健壮和可扩展的策略，用于IPSC来源的CMS的大规模生产。在搅拌槽或波浪生物反应器中，将小鼠IPSCs培养成聚集体。评价了不同水动力环境促进的溶解氧和机械力对CM分化的影响。在搅拌槽生物反应器中，将低氧培养(4%O2张力)与间歇搅拌相结合，与常氧(20%O2张力)和连续搅拌培养相比，CM产量提高了约1000倍。此外，我们首次展示了波诱导搅拌使IPSC能够以更快的动力学和更高的产率(60CMS/输入IPSC)分化为CMS。在11天的分化方案中，产生了临床上相关数量的CMS(2.3 × 10 9 CMS/1个L)，并且CMS具有典型的心肌肌节结构、钙瞬变、电生理特征和药物敏感性。这项工作描述了小鼠IPSC向可伸缩心肌细胞分化方面的重大进展，为实施这一策略大规模生产人类同类细胞并将其用于心脏修复和心血管研究铺平了道路。本文的在线版本(doi：10.1007/s12015-0149533-0)包含补充材料，可供授权用户使用。
Cardiomyocytes (CMs) derived from induced pluripotent stem cells (iPSCs) hold great promise for patient-specific disease modeling, drug screening and cell therapy. However, existing protocols for CM differentiation of iPSCs besides being highly dependent on the application of expensive growth factors show low reproducibility and scalability. The aim of this work was to develop a robust and scalable strategy for mass production of iPSC-derived CMs by designing a bioreactor protocol that ensures a hypoxic and mechanical environment. Murine iPSCs were cultivated as aggregates in either stirred tank or WAVE bioreactors. The effect of dissolved oxygen and mechanical forces, promoted by different hydrodynamic environments, on CM differentiation was evaluated. Combining a hypoxia culture (4 % O2 tension) with an intermittent agitation profile in stirred tank bioreactors resulted in an improvement of about 1000-fold in CM yields when compared to normoxic (20 % O2 tension) and continuously agitated cultures. Additionally, we showed for the first time that wave-induced agitation enables the differentiation of iPSCs towards CMs at faster kinetics and with higher yields (60 CMs/input iPSC). In an 11-day differentiation protocol, clinically relevant numbers of CMs (2.3 × 109 CMs/1 L) were produced, and CMs exhibited typical cardiac sarcomeric structures, calcium transients, electrophysiological profiles and drug responsiveness. This work describes significant advances towards scalable cardiomyocyte differentiation of murine iPSC, paving the way for the implementation of this strategy for mass production of their human counterparts and their use for cardiac repair and cardiovascular research. The online version of this article (doi:10.1007/s12015-014-9533-0) contains supplementary material, which is available to authorized users.
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影响因子：
3.8
作者：
Bauwens, C;Yin, T;Zandstra, PW
通讯作者：
Zandstra, PW
影响因子：
10.8
作者：
Halbach, Marcel;Peinkofer, Gabriel;Mueller-Ehmsen, Jochen
通讯作者：
Mueller-Ehmsen, Jochen
影响因子：
--
作者：
Matsumoto, Takuya;Yung, Yu Ching;Mooney, David J.
通讯作者：
Mooney, David J.
DOI：
10.1089/clo.2006.8.117
发表时间：
2006-06-01
期刊：
CLONING AND STEM CELLS
影响因子：
--
作者：
Gibbons, J.;Hewitt, E.;Gardner, D. K.
通讯作者：
Gardner, D. K.
影响因子：
64.8
作者：
Laflamme, Michael A.;Murry, Charles E.
通讯作者：
Murry, Charles E.