Thermoresponsive Worms for Expansion and Release of Human Embryonic Stem Cells

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• 作者：Xiaoli Chen ; Andrew B. J. Prowse ; Zhongfan Jia ; Helena Tellier ; Trent P. Munro ; Peter P. Gray ; Michael J. Monteiro
• 刊名：Biomacromolecules
• 出版年：2014
• 出版时间：March 10, 2014
• 年：2014
• 卷：15
• 期：3
• 页码：844-855
• 全文大小：648K
• 年卷期：v.15,no.3(March 10, 2014)
• ISSN：1526-4602

文摘

The development of robust suspension cultures of human embryonic stem cells (hESCs) without the use of cell membrane disrupting enzymes or inhibitors is critical for future clinical applications in regenerative medicine. We have achieved this by using long, flexible, and thermoresponsive polymer worms decorated with a recombinant vitronectin subdomain that bridge hESCs, aiding in hESC鈥檚 natural ability to form embryoid bodies (EBs) and satisfying their inherent requirement for cell鈥揷ell and cell鈥揺xtracellular matrix contact. When the EBs reached an optimal upper size where cytokine and nutrient penetration becomes limiting, these long and flexible polymer worms facilitated EB breakdown via a temperature shift from 37 to 25 掳C. The thermoresponsive nature of the worms enabled a cyclical dissociation and propagation of the cells. Repeating the process for three cycles (over eighteen days) provided a >30-fold expansion in cell number while maintaining pluripotency, thereby providing a simple, nondestructive process for the 3D expansion of hESC.