Critical microcarrier properties affecting the expansion of undifferentiated human embryonic stem cells
Critical microcarrier properties affecting the expansion of undifferentiated human embryonic stem cells
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DOI：
10.1016/j.scr.2011.04.007
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发表时间：
2011-09-01
影响因子：
1.2
通讯作者：
Oh, Steve Kah Weng
中科院分区：
文献类型：
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作者：
Chen, Allen Kuan-Liang;Chen, Xiaoli;Oh, Steve Kah Weng
各种微载体可用于扩增人胚胎干细胞(HESC)以用于细胞治疗应用。本研究考察了10种微载体对人胚胎干细胞贴壁效率、生长和多能性的影响。在未包被的微载体上观察到很高的附着效率，但在连续传代过程中会出现细胞生长不良和/或逐渐丧失多能性。用Matrigel包覆微载体可产生更高的细胞产率和至少三代的稳定的多潜能状态。带正电的圆柱形纤维素微载体(DE52、DE53和QA52)和大(190微米)带正电的球形微载体(Cytodex 1)具有较高的细胞扩增潜力和多能性。使用直径较小的球形(65微米和10微米)或大孔微珠可获得较低的细胞产量。代替Matrigel，层粘连蛋白包裹的微载体(DE53和Cytodex 1)能够支持HES-2和HES-3细胞系的长期增殖和多能性。在旋转瓶中，早期抗剪切的HES-2细胞株在Matrigel(84%Tra-1-60，1.43×10(6)cell/ml)和层粘连蛋白(74%Tra-1-60，1.37×10(6)cell/ml)包被的微载体上培养时，获得了相似的细胞生长和多潜能标记的表达。相反，与Matrigel包被的微载体相比，HES-3的细胞产量、存活率和层粘连蛋白的多能标志物都有所降低，这可能是由于剪切敏感性所致。传统的用于哺乳动物细胞增殖的微载体不适合hESC的长期增殖。基质或层粘连蛋白涂层对于hESC在各种微载体上的长期稳定繁殖是必不可少的。(C)2011爱思唯尔B.V.保留所有权利。
A variety of microcarriers may be used for the expansion of human embryonic stem cells (hESC) for cell therapy applications. This study investigated the effects of 10 types of microcarriers on hESC attachment efficiency, growth and pluripotency. High attachment efficiency was observed on uncoated microcarriers, however poor cell growth and/or gradual loss of pluripotency occurred during continuous passaging. Coating of the microcarriers with Matrigel resulted in higher cell yields and stable pluripotent states for at least three passages. Positively charged cylindrical cellulose microcarriers (DE52, DE53 and QA52) and large (190 mu m) positively charged spherical microcarriers (Cytodex 1) exhibited high cell expansion potential and levels of pluripotency. Lower cell yields were obtained using smaller diameter spherical (65 mu m and 10 mu m) or macroporous beads. Instead of Matrigel, laminin coated microcarriers (DE53 and Cytodex 1) are capable of supporting the long term propagation and pluripotency of HES-2 and HES-3 cell lines. HES-2 cell line which was shown earlier to be shear resistant achieved similar cell growth and expression of pluripotent markers when cultured on both Matrigel (84% Tra-1-60, 1.43 x 10(6) cells/ml) and laminin (74% Tra-1-60, 1.37 x 10(6) cells/ml) coated microcarriers in spinner flasks. In contrast, HES-3 exhibited a decrease in cell yield, viability and pluripotent markers on laminin as compared with Matrigel coated microcarriers possibly due to shear sensitivity. Conventional microcarriers intended for propagation of mammalian cells are not suitable for long term propagation of hESC. Matrigel or laminin coating is essential for stable long term propagation of hESC on a variety of microcarriers. (C) 2011 Elsevier B.V. All rights reserved.