Abstract：
Fluorinated ethylene propylene (FEP) vessels are of significant interest for therapeutic cell biomanufacturing applications due to their chemical inertness, hydrophobic surface, and high oxygen permeability. However, these properties also limit the adhesion and survival of anchorage-dependent cells. Here, we develop novel plasma polymer coatings to modify FEP surfaces, enhancing the adhesion and expansion of human mesenchymal stromal cells (hMSCs). Similar to commercially available tissue culture polystyrene vessels, oxygen-rich or nitrogen-rich surface chemistries can be achieved using this approach. While steam sterilization increased the roughness of the coatings and altered the surface chemistry, the overall wettability and oxygen or nitrogen-rich nature of the coatings were maintained. In the absence of proteins during initial cell attachment, cells adhered to surfaces even in the presence of chelators, whereas adhesion was abrogated with chelator in a protein-containing medium, suggesting that integrin-mediated adhesion predominates over physicochemical tethering in normal protein-containing cell seeding conditions. Albumin adsorption was more elevated on nitrogen-rich coatings compared to the oxygen-rich coatings, which was correlated with a higher extent of hMSC expansion after 3 days. Both the oxygen and nitrogen-rich coatings significantly improved hMSC adhesion and expansion compared to untreated FEP. FEP surfaces with nitrogen-rich coatings were practically equivalent to commercially available standard tissue culture-treated polystyrene surfaces in terms of hMSC yields. Plasma polymer coatings show significant promise in expanding the potential usage of FEP-based culture vessels for cell therapy applications.
摘要：
氟化乙烯丙烯（FEP）容器由于其化学惰性而对治疗细胞生物制造应用具有重要意义，疏水表面，和高氧气渗透性。然而,这些特性也限制了贴壁依赖性细胞的粘附和存活。这里,我们开发了新型等离子体聚合物涂层来改性FEP表面，增强人间充质基质细胞（hMSCs）的粘附和扩增。与市售组织培养聚苯乙烯容器相似，富氧或富氮的表面化学可以使用这种方法来实现。虽然蒸汽灭菌增加了涂层的粗糙度并改变了表面化学，涂层的总体润湿性和富氧或富氮性质得以保持。在最初的细胞附着过程中没有蛋白质的情况下，即使在存在螯合剂的情况下，细胞也会粘附在表面，而在含蛋白质的培养基中用螯合剂消除粘附，表明在正常的含蛋白质的细胞接种条件下，整合素介导的粘附优于物理化学连接。与富氧涂层相比，富氮涂层上的白蛋白吸附更高，这与3天后更高的hMSC扩增程度相关。与未处理的FEP相比,富氧和富氮涂层都显著改善了hMSC的粘附性和膨胀。就hMSC产率而言，具有富氮涂层的FEP表面实际上与市售标准组织培养处理的聚苯乙烯表面相当。等离子体聚合物涂层在扩大基于FEP的培养容器用于细胞治疗应用的潜在用途方面显示出重大前景。
