High-Throughput Discovery of Targeted, Minimally Complex Peptide Surfaces for Human Pluripotent Stem Cell Culture.
High-Throughput Discovery of Targeted, Minimally Complex Peptide Surfaces for Human Pluripotent Stem Cell Culture.
复制标题
用于人类多能干细胞培养的靶向、最小复杂肽表面的高通量发现。
DOI：
10.1021/acsbiomaterials.0c01462
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发表时间：
2021
影响因子：
5.8
通讯作者：
Schaffer,DavidV
中科院分区：
文献类型：
--
作者：
Ramasubramanian,Anusuya;Muckom,Riya;Sugnaux,Caroline;Fuentes,Christina;Ekerdt,BarbaraL;Clark,DouglasS;Healy,KevinE;Schaffer,DavidV
人类多能干细胞具有无限的能力来产生用于再生医学的治疗相关细胞。然而，利用这些细胞在临床上，可扩展的培养系统，激活定义的受体和信号通路，以维持干细胞的自我更新是必需的;和合成材料提供了相当大的承诺，以满足这些needs.De novodevelopment的材料，目标新的途径一直受到阻碍的关键受体相互作用维持多能性的有限理解。在这里，我们通过无偏的、基于文库的淘选策略鉴定了人多能干细胞（hPSC）层粘连蛋白受体和多能性调节因子α6-整联蛋白的肽激动剂。粘附的生物物理表征表明，所鉴定的肽通过α6-整联蛋白结合hPSC，其解离常数与层粘连蛋白相似，为亚μM。通过利用高通量微培养平台，我们开发了预测指南，用于以最佳化学计量将这些整合素靶向肽与经典结合基序一起呈现，以产生新生培养表面。最后，当以自组装单层呈现时，预测的肽组合支持hPSC扩增，突出了无偏筛选如何加速靶向生物材料的发现。
Human pluripotent stem cells harbor an unlimited capacity to generate therapeutically relevant cells for applications in regenerative medicine. However, to utilize these cells in the clinic, scalable culture systems that activate defined receptors and signaling pathways to sustain stem cell self-renewal are required; and synthetic materials offer considerable promise to meet these needs.De novodevelopment of materials that target novel pathways has been stymied by a limited understanding of critical receptor interactions maintaining pluripotency. Here, we identify peptide agonists for the human pluripotent stem cell (hPSC) laminin receptor and pluripotency regulator, α6-integrin, through unbiased, library-based panning strategies. Biophysical characterization of adhesion suggests that identified peptides bind hPSCs through α6-integrin with sub-μM dissociation constants similar to laminin. By harnessing a high-throughput microculture platform, we developed predictive guidelines for presenting these integrin-targeting peptides alongside canonical binding motifs at optimal stoichiometries to generate nascent culture surfaces. Finally, when presented as self-assembled monolayers, predicted peptide combinations supported hPSC expansion, highlighting how unbiased screens can accelerate the discovery of targeted biomaterials.