Department of Biomedical Engineering, University of Wisconsin-Madison, 1550 Engineering Dr., 53706, USA.
Department of Orthopedics and Rehabilitation, University of Wisconsin-Madison, 1685 Highland Ave., 53705, USA.
Macromol Biosci. 2019 Feb;19(2):e1800299. doi: 10.1002/mabi.201800299. Epub 2018 Nov 22.
Mesenchymal stem cells (MSC), also called marrow stromal cells, are adult cells that have attracted interest for their potential uses in therapeutic applications. There is a pressing need for scalable culture systems due to the large number of cells needed for clinical treatments. Here, a tailorable thin polymer coating-poly(poly(ethylene glycol) methyl ether methacrylate-ran-vinyl dimethyl azlactone-ran-glycidyl methacrylate) [P(PEGMEMA-r-VDM-r-GMA); PVG]-to the surface of commercially available polystyrene and glass microcarriers to create chemically defined surfaces for large-scale cell expansion is applied. These chemically defined microcarriers create a reproducible surface that does not rely on the adsorption of xenogenic serum proteins to mediate cell adhesion. Specifically, this coating method anchors PVG copolymer through ring opening nucleophilic attack by amine residues on poly-l-lysine that is pre-adsorbed to the surface of microcarriers. Importantly, this anchoring reaction preserves the monomer VDM reactivity for subsequent functionalization with an integrin-specific Arg-Gly-Asp peptide to enable cell adhesion and expansion via a one-step reaction in aqueous media. MSCs cultured on PVG-coated microcarriers achieve sixfold expansion-similar to the expansion achieved on PS microcarriers-and retain their ability to differentiate after harvesting.
间充质干细胞（MSC），也称为骨髓基质细胞，是一种成体细胞，因其在治疗应用中的潜在用途而受到关注。由于临床治疗需要大量的细胞，因此迫切需要可扩展的培养系统。在这里，我们将一种可定制的薄聚合物涂层-聚（聚（乙二醇）甲基醚甲基丙烯酸酯-共聚-乙烯基二甲基偶氮内酯-共聚-甲基丙烯酸缩水甘油酯）[P（PEGMEMA-r-VDM-r-GMA）；PVG]应用于市售聚苯乙烯和玻璃微载体的表面，以创建用于大规模细胞扩增的化学定义表面。这些化学定义的微载体创造了一种可重复的表面，不依赖于异种血清蛋白的吸附来介导细胞黏附。具体来说，这种涂层方法通过预先吸附在微载体表面的聚赖氨酸上的胺残基的开环亲核攻击来固定 PVG 共聚物。重要的是，这种固定反应保留了单体 VDM 的反应性，可用于随后与整联蛋白特异性的 Arg-Gly-Asp 肽进行功能化，从而通过在水性介质中的一步反应实现细胞黏附和扩增。在 PVG 涂层微载体上培养的 MSC 可实现六倍的扩增-与在 PS 微载体上实现的扩增相似-并在收获后保留其分化能力。
