Schmitt Samantha K, Xie Angela W, Ghassemi Raha M, Trebatoski David J, Murphy William L, Gopalan Padma
Department of Materials Science and Engineering, University of Wisconsin-Madison, Madison, WI, 53706, USA.
Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, 53706, USA.
Adv Healthc Mater. 2015 Jul 15;4(10):1555-64. doi: 10.1002/adhm.201500191. Epub 2015 May 20.
Human mesenchymal stem cells (hMSCs) are a widely available and clinically relevant cell type with a host of applications in regenerative medicine. Current clinical expansion methods can lead to selective changes in hMSC phenotype potentially resulting from relatively undefined cell culture surfaces. Chemically defined synthetic surfaces can aid in understanding the influence of cell-material interactions on stem cell behavior. Here, a thin copolymer coating for hMSC culture on plastic substrates is developed. The random copolymer is synthesized by living free radical polymerization and characterized in solution before application to the substrate, ensuring a homogeneous coating and limiting the sample-to-sample variations. The ability to coat multiple substrate types and cover large surface areas is reported. Arg-Gly-Asp-containing peptides are incorporated into the coating under aqueous conditions via their lysine or cysteine side chains, resulting in amide and thioester linkages, respectively. Stability studies show amide linkages to be stable and thioester linkages to be labile under standard serum-containing culture conditions. In addition, chemically defined passaging of hMSCs using only ethylenediaminetetraacetic acid on polystyrene dishes is shown. After passage, the hMSCs can be seeded back onto the same plate, indicating potential reusability of the coating.
人间充质干细胞（hMSCs）是一种广泛可用且与临床相关的细胞类型，在再生医学中有诸多应用。当前的临床扩增方法可能会导致hMSC表型发生选择性变化，这可能是由相对不明确的细胞培养表面引起的。化学定义的合成表面有助于理解细胞 - 材料相互作用对干细胞行为的影响。在此，开发了一种用于在塑料基质上培养hMSC的薄共聚物涂层。该无规共聚物通过活性自由基聚合合成，并在应用于基质之前在溶液中进行表征，以确保涂层均匀并限制样品间的差异。报告了涂覆多种基质类型和覆盖大面积的能力。含精氨酸 - 甘氨酸 - 天冬氨酸的肽通过其赖氨酸或半胱氨酸侧链在水性条件下掺入涂层中，分别形成酰胺键和硫酯键。稳定性研究表明，在含标准血清的培养条件下，酰胺键稳定，硫酯键不稳定。此外，还展示了在聚苯乙烯培养皿上仅使用乙二胺四乙酸对hMSC进行化学定义传代的方法。传代后，hMSC可以重新接种到同一培养皿上，表明该涂层具有潜在的可重复使用性。
