Three-dimensional culture systems for the expansion of pluripotent embryonic stem cells.
Three-dimensional culture systems for the expansion of pluripotent embryonic stem cells.
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DOI：
10.1002/bit.22850
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发表时间：
2010-11-01
影响因子：
3.8
通讯作者：
Welham, Melanie J.
中科院分区：
文献类型：
--
作者：
Storm, Michael P.;Orchard, Craig B.;Bone, Heather K.;Chaudhuri, Julian B.;Welham, Melanie J.
小鼠胚胎干细胞（ESC）系和最近的人类ESC系已成为研究早期哺乳动物发育的宝贵工具。对ESC及其分化后代在药物发现和作为潜在治疗剂中的日益增加的兴趣突出了当前二维（2D）静态培养技术不足以进行大规模生产的事实。在三维（3D）搅拌系统中培养哺乳动物细胞已被证明克服了2D的许多限制，因此可能对ESC增殖有效。使用小鼠胚胎干细胞作为我们的初始模型，我们研究了不同3D培养环境对多能胚胎干细胞扩增的有效性。采用Solohill胶原蛋白、Solohill FACT和Cultispher-S微载体，并与搅拌生物反应器结合使用。初始接种参数，包括细胞数量和搅拌条件，被认为是至关重要的，在促进附着到微载体和最大限度地减少所形成的聚集体的大小。虽然所有微载体都支持未分化mESC的生长，但Cultispher-S的性能优于Solohill微载体。当在Cultispher-S微载体上连续传代培养时，mESC保持其多能性，这通过自我更新、多能性标志物的表达和经历多谱系分化的能力来证明。当将这些优化的条件应用于未断奶的人ESC时，Cultispher-S微载体支持hESC的生长，所述hESC保留多能性标志物（包括SSEA 4、Tra-1-60、NANOG和OCT-4）的表达。我们的研究强调了初始接种参数优化的重要性，并提供了概念验证数据，证明了微载体和生物反应器用于扩增hESC的实用性。Biotechnol. Bioeng. 2010;107：683-695。© 2010 Wiley Periodicals，Inc.
Mouse embryonic stem cell (ESC) lines, and more recently human ESC lines, have become valuable tools for studying early mammalian development. Increasing interest in ESCs and their differentiated progeny in drug discovery and as potential therapeutic agents has highlighted the fact that current two-dimensional (2D) static culturing techniques are inadequate for large-scale production. The culture of mammalian cells in three-dimensional (3D) agitated systems has been shown to overcome many of the restrictions of 2D and is therefore likely to be effective for ESC proliferation. Using murine ESCs as our initial model, we investigated the effectiveness of different 3D culture environments for the expansion of pluripotent ESCs. Solohill Collagen, Solohill FACT, and Cultispher-S microcarriers were employed and used in conjunction with stirred bioreactors. Initial seeding parameters, including cell number and agitation conditions, were found to be critical in promoting attachment to microcarriers and minimizing the size of aggregates formed. While all microcarriers supported the growth of undifferentiated mESCs, Cultispher-S out-performed the Solohill microcarriers. When cultured for successive passages on Cultispher-S microcarriers, mESCs maintained their pluripotency, demonstrated by self-renewal, expression of pluripotency markers and the ability to undergo multi-lineage differentiation. When these optimized conditions were applied to unweaned human ESCs, Cultispher-S microcarriers supported the growth of hESCs that retained expression of pluripotency markers including SSEA4, Tra-1–60, NANOG, and OCT-4. Our study highlights the importance of optimization of initial seeding parameters and provides proof-of-concept data demonstrating the utility of microcarriers and bioreactors for the expansion of hESCs. Biotechnol. Bioeng. 2010;107:683–695. © 2010 Wiley Periodicals, Inc.
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DOI：
10.1590/s0100-879x2009000600007
发表时间：
2009-06-01
影响因子：
2.3
作者：
Fernandes, A.M.;Marinho, P.A.N.;Rehen, S.K.
通讯作者：
Rehen, S.K.
影响因子：
3.9
作者：
Welham, M. J.;Storm, M. P.;Bone, H. K.
通讯作者：
Bone, H. K.
影响因子：
64.8
作者：
SMITH, AG;HEATH, JK;ROGERS, D
通讯作者：
ROGERS, D
影响因子：
2.9
作者：
Nie, Ying;Bergendahl, Veit;Hei, Derek J.;Jones, Jeffrey M.;Palecek, Sean P.
通讯作者：
Palecek, Sean P.
影响因子：
4.1
作者：
Phillips, Blaine Wesley;Horne, Rachel;Crook, Jeremy Micah
通讯作者：
Crook, Jeremy Micah