Synthetic, Chemically Defined Polymer-Coated Microcarriers for the Expansion of Human Mesenchymal Stem Cells
Synthetic, Chemically Defined Polymer-Coated Microcarriers for the Expansion of Human Mesenchymal Stem Cells
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DOI：
10.1002/mabi.201800299
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发表时间：
2019-02-01
影响因子：
4.6
通讯作者：
Gopalan, Padma
中科院分区：
文献类型：
--
作者：
Krutty, John D.;Dias, Andrew D.;Gopalan, Padma
间充质干细胞（MSC），也称为骨髓基质细胞，是成体细胞，因其在治疗应用中的潜在用途而引起人们的兴趣。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，将可定制的薄聚合物涂层-聚（聚（乙二醇）甲基醚甲基丙烯酸酯-ran-乙烯基二甲基氮杂内酯-ran-甲基丙烯酸缩水甘油酯）[P（PEGMEMA-r-VDM-r-GMA）; PVG]-涂在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。这些化学成分确定的微载体产生了一个可再现的表面，不依赖于异种血清蛋白的吸附来介导细胞粘附。具体地，该涂覆方法通过胺残基对预吸附到微载体表面的聚-1-赖氨酸的开环亲核攻击来锚定PVG共聚物。重要的是，这种锚定反应保留了单体VDM的反应性，用于随后用整联蛋白特异性Arg-Gly-Asp肽进行官能化，以通过在水性介质中的一步反应实现细胞粘附和扩增。在PVG包被的微载体上培养的MSC实现了六倍的扩增-类似于在PS微载体上实现的扩增-并且在收获后保留了它们的分化能力。
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.