N-Isopropylacrylamide-Based Thermoresponsive Polyelectrolyte Multilayer Films for Human Mesenchymal Stem Cell Expansion
N-Isopropylacrylamide-Based Thermoresponsive Polyelectrolyte Multilayer Films for Human Mesenchymal Stem Cell Expansion
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DOI：
10.1002/btpr.471
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发表时间：
2010-11-01
影响因子：
2.9
通讯作者：
Ma, Teng
中科院分区：
文献类型：
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作者：
Liao, Tianqing;Moussallem, Maroun D.;Ma, Teng
人骨髓间充质干细胞(HMSCs)是一种来源于成人骨髓的集落形成单位成纤维细胞(CFU-F)，在许多基于细胞的组织工程应用中具有巨大的潜力。然而，随着它们在培养中的扩张，它们的可塑性逐渐减弱，这限制了它们的治疗潜力。在本研究中，我们使用N-异丙基丙烯酰胺(NIPAM)基热响应性聚电解质多层膜(N-PEMU)作为培养基质来支持hMSC的扩增，并评价了它们对细胞特性的影响。N-PEMU膜是通过与带电单体、带正电的烯丙胺盐酸盐(PAH)或带负电的苯乙烯磺酸(PSS)共聚的温敏性单体逐层吸附制备的，并与胎牛血清(FBS)涂层表面进行了比较。表面电荷可以改变细胞外基质(ECM)结构，进而调节hMSC的黏附、增殖、整合素表达、脱离和集落形成能力。带正电的热响应表面在与对照表面相当的范围内改善了细胞的粘附性和生长，同时保持了显著更高的CFU-F形成能力。免疫染色和Western印迹结果表明，细胞在正电荷表面的黏附和生长的改善是由于纤维连接蛋白等ECM蛋白在正电荷表面的黏附增加所致。这些结果表明，逐层方法是一种不需要酶处理就可以形成基于PNIPAM的热响应表面的有效方法，可以用于hMSC的生长和去除。结果还表明，表面电荷调节ECM的黏附，这不仅影响细胞的黏附，而且还影响CFU的形成能力和多系分化潜能。(C)2010年美国化学工程师学会生物技术研究所。程序，26：1705-1713,2010
Human mesenchymal stem cells (hMSCs) are colony-forming unit fibroblasts (CFU-F) derived from adult bone marrow and have significant potential for many cell-based tissue-engineering applications. Their therapeutic potential, however, is restricted by their diminishing plasticity as they are expanded in culture. In this study, we used N-isopropylacrylamide (NIPAM)-based thermoresponsive polyelectrolyte multilayer (N-PEMU) films as culture substrates to support hMSC expansion and evaluated their effects on cell properties. The N-PEMU films were made via layer-by-layer adsorption of thermoresponsive monomers copolymerized with charged monomers, positively charged allylamine hydrochloride (PAH), or negatively charged styrene sulfonic acid (PSS) and compared to fetal bovine serum (FBS) coated surfaces. Surface charges were shown to alter the extracellular matrix (ECM) structure and subsequently regulate hMSC responses including adhesion, proliferation, integrin expression, detachment, and colony forming ability. The positively charged thermal responsive surfaces improved cell adhesion and growth in a range comparable to control surfaces while maintaining significantly higher CFU-F forming ability. Immunostaining and Western blot results indicate that the improved cell adhesion and growth on the positively charged surfaces resulted from the elevated adhesion of ECM proteins such as fibronectin on the positively charge surfaces. These results demonstrate that the layer-by-layer approach is an efficient way to form PNIPAM-based thermal responsive surfaces for hMSC growth and removal without enzymatic treatment. The results also show that surface charge regulates ECM adhesion, which in turn influences not only cell adhesion but also CFU-forming ability and their multi-lineage differentiation potential. (C) 2010 American Institute of Chemical Engineers Biotechnol. Prog., 26: 1705-1713, 2010