A synthetic, xeno-free peptide surface for expansion and directed differentiation of human induced pluripotent stem cells.
A synthetic, xeno-free peptide surface for expansion and directed differentiation of human induced pluripotent stem cells.
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DOI：
10.1371/journal.pone.0050880
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Ye K
中科院分区：
文献类型：
--
作者：
Jin S;Yao H;Weber JL;Melkoumian ZK;Ye K
人类诱导多能干细胞有潜力成为细胞替代疗法的无限细胞来源。然而，这种潜力的实现取决于培养方法的可用性，这些方法是稳健的，可扩展的，并使用化学定义的材料。尽管hiPSC技术取得了重大进展，但hiPSC的扩增依赖于使用动物源性细胞外基质提取物，如Matrigel，这引起了对使用这些产品的安全性担忧。在这项工作中，我们研究了在化学成分确定的无异种合成肽底物（即Corning Synthemax® Surface）上扩增和分化hiPSC的可行性。我们证明了Synthemax表面支持hiPSC的附着、铺展和增殖，以及hiPSC的谱系特异性分化。与在Matrigel™上生长的细胞相比，在Synthemax Surfaces上生长的hiPSC集落表现出更少的扩散和更紧凑的形态。在Synthemax表面上生长的hiPSC的细胞骨架表征揭示了在细胞-细胞界面中形成更致密的肌动蛋白丝。在Synthemax表面上生长的hiPSC中也观察到黏着斑蛋白的下调和zyxin表达的上调。细胞-ECM相互作用的进一步检查显示，在Synthemax表面上生长的hiPSC主要利用αvβ5整联蛋白介导与基质的附着，而多种整联蛋白参与细胞与基质胶的附着。最后，hiPSC可以在Synthemax表面上保持未分化超过十代。这些研究提供了一种使用合成肽工程化表面作为底物扩增hiPSC的新方法，以避免动物源性材料的污染和批间变异性的潜在风险。
Human induced pluripotent stem cells have the potential to become an unlimited cell source for cell replacement therapy. The realization of this potential, however, depends on the availability of culture methods that are robust, scalable, and use chemically defined materials. Despite significant advances in hiPSC technologies, the expansion of hiPSCs relies upon the use of animal-derived extracellular matrix extracts, such as Matrigel, which raises safety concerns over the use of these products. In this work, we investigated the feasibility of expanding and differentiating hiPSCs on a chemically defined, xeno-free synthetic peptide substrate, i.e. Corning Synthemax® Surface. We demonstrated that the Synthemax Surface supports the attachment, spreading, and proliferation of hiPSCs, as well as hiPSCs’ lineage-specific differentiation. hiPSCs colonies grown on Synthemax Surfaces exhibit less spread and more compact morphology compared to cells grown on Matrigel™. The cytoskeleton characterization of hiPSCs grown on the Synthemax Surface revealed formation of denser actin filaments in the cell-cell interface. The down-regulation of vinculin and up-regulation of zyxin expression were also observed in hiPSCs grown on the Synthemax Surface. Further examination of cell-ECM interaction revealed that hiPSCs grown on the Synthemax Surface primarily utilize αvβ5 integrins to mediate attachment to the substrate, whereas multiple integrins are involved in cell attachment to Matrigel. Finally, hiPSCs can be maintained undifferentiated on the Synthemax Surface for more than ten passages. These studies provide a novel approach for expansion of hiPSCs using synthetic peptide engineered surface as a substrate to avoid a potential risk of contamination and lot-to-lot variability with animal derived materials.
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影响因子：
14
作者：
Dalby, MJ;Riehle, MO;Curtis, ASG
通讯作者：
Curtis, ASG
DOI：
10.1083/jcb.200204153
发表时间：
2002-11-25
期刊：
The Journal of cell biology
影响因子：
--
作者：
Galbraith CG;Yamada KM;Sheetz MP
通讯作者：
Sheetz MP
影响因子：
21.3
作者：
Balaban, NQ;Schwarz, US;Geiger, B
通讯作者：
Geiger, B
影响因子：
7.5
作者：
Bresnick, AR
通讯作者：
Bresnick, AR
影响因子：
4.8
作者：
Besser, D
通讯作者：
Besser, D