Mesenchymal stem cell therapy has suffered from wide variability in clinical efficacy, largely due to heterogeneous starting cell populations and large-scale cell death during and after implantation. Optimizing the manufacturing process has led to reproducible cell populations that can be cryopreserved for clinical applications. Nevertheless, ensuring a reproducible cell state that persists after cryopreservation remains a significant challenge, and is necessary to ensure reproducible clinical outcomes. Here we demonstrate how matrix-conjugated hydrogel cell culture materials can normalize a population of induced pluripotent stem cell derived mesenchymal stem cells (iPSC-MSCs) to display a defined secretory profile that promotes enhanced neovascularization in vitro and in vivo. Using a protein-conjugated biomaterials screen we identified two conditions—1 kPa collagen and 10 kPa fibronectin coated polyacrylamide gels—that promote reproducible secretion of pro-angiogenic and immunomodulatory cytokines from iPSC-MSCs that enhance tubulogenesis of endothelial cells in Geltrex and neovascularization in chick chorioallantoic membranes. Using defined culture substrates alone, we demonstrate maintenance of secretory activity after cryopreservation for the first time. This advance provides a simple and scalable approach for cell engineering and subsequent manufacturing, toward normalizing and priming a desired cell activity for clinical regenerative medicine.

中文翻译：

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生物材料在诱导多能干细胞衍生的间充质基质细胞中定向激活低温治疗性分泌组

间充质干细胞疗法的临床疗效差异很大，这主要是由于起始细胞群的异质性以及植入期间和植入后的大规模细胞死亡。优化制造过程已经产生了可重现的细胞群，这些细胞群可以冷冻保存用于临床应用。然而，确保冷冻保存后持续存在的可重现细胞状态仍然是一项重大挑战，并且对于确保可重现的临床结果是必要的。在这里，我们展示了基质结合的水凝胶细胞培养材料如何使诱导多能干细胞衍生的间充质干细胞 (iPSC-MSC) 群体正常化，以显示明确的分泌特征，从而促进体外和体内新血管形成。使用蛋白质结合的生物材料筛选，我们确定了两种条件——1 kPa 胶原蛋白和 10 kPa 纤连蛋白涂层聚丙烯酰胺凝胶——促进 iPSC-MSC 可重复分泌促血管生成和免疫调节细胞因子，增强 Geltrex 中内皮细胞的小管生成和小鸡的新血管形成绒毛尿囊膜。单独使用确定的培养基质，我们首次证明了冷冻保存后分泌活动的维持。这一进展为细胞工程和后续制造提供了一种简单且可扩展的方法，以实现临床再生医学所需细胞活性的正常化和启动。