Future Industries Institute, University of South Australia, Mawson Lakes, Adelaide, SA 5095, Australia; Cooperative Research Centre for Cell Therapy Manufacturing (CRC-CTM), Mawson Lakes, SA 5095, Australia.
Future Industries Institute, University of South Australia, Mawson Lakes, Adelaide, SA 5095, Australia; Cooperative Research Centre for Cell Therapy Manufacturing (CRC-CTM), Mawson Lakes, SA 5095, Australia.
Colloids Surf B Biointerfaces. 2018 Jan 1;161:261-268. doi: 10.1016/j.colsurfb.2017.10.065. Epub 2017 Oct 26.
Human Mesenchymal Stem cells (hMSCs) are becoming a major focus in biomedical fields. Application of in vitro expanded hMSCs to treat numerous ailments has led to a commercial emphasis on improving hMSC growth ex vivo. Production of substrate independent, novel thin films is one potential tool for production of commercial viable hMSC expansion. Plasma polymerization allow controlled chemical optimisation of large scale surface areas in a substrate independent manner. Previous study shown that plasma polymerized Furfuryl Methacrylate (ppFMA) surfaces allowed primary fibroblast cells adhesion and proliferation. However, under some deposition conditions, particle aggregates formation was observed. These aggregates had the effect of disrupting cell attachment, despite being chemically indistinguishable from the underlying surface. Herein, hMSCs were cultured on ppFMA surfaces to determine their suitability for stem cell culture and observe the effect of particle aggregates on hMSC attachment and growth. Both metabolic and DNA quantification assays showed that surfaces with particle aggregates had lower numbers of attached cells and slower growth. Uniform surfaces without aggregates showed higher cell attachment and growth levels, which were comparable to Thermanox. Phenotypic analysis showed that there was no change to hMSCs phenotype after 7 & 14days of culture on uniform ppFMA surface. Further investigation using time-lapse image analysis indicated that particle aggregates reduced cell attachment by presenting a physically weak boundary layer, which was damaged by intracellular tension during cell spreading. ppFMA surface can provide a stable substrate independent hMSCs expansion interface that could be applied to larger scale bioreactors, beads or scaffolds.
人骨髓间充质干细胞(hMSCs)正在成为生物医学领域的一个主要焦点。体外扩增的 hMSCs 应用于治疗多种疾病,这导致了商业界对 ex vivo 提高 hMSC 生长的重视。生产独立于基质的新型薄膜是生产商业上可行的 hMSC 扩展的潜在工具之一。等离子体聚合允许以独立于基质的方式对大表面积进行受控的化学优化。先前的研究表明,等离子体聚合的糠基甲基丙烯酸酯(ppFMA)表面允许原代成纤维细胞的黏附和增殖。然而,在某些沉积条件下,观察到了颗粒聚集体的形成。尽管这些聚集体在化学上与底层表面没有区别,但它们会破坏细胞附着。在此,将 hMSCs 培养在 ppFMA 表面上,以确定其在干细胞培养中的适用性,并观察颗粒聚集体对 hMSC 附着和生长的影响。代谢和 DNA 定量分析均表明,具有颗粒聚集体的表面附着的细胞数量较少,生长速度较慢。无聚集体的均匀表面显示出更高的细胞附着和生长水平,与 Thermanox 相当。表型分析表明,在均匀的 ppFMA 表面上培养 7 天和 14 天后,hMSCs 的表型没有变化。使用延时图像分析的进一步研究表明,颗粒聚集体通过呈现物理上较弱的边界层来减少细胞附着,该边界层在细胞扩展过程中会被细胞内张力破坏。ppFMA 表面可为独立于基质的 hMSCs 扩展提供稳定的界面,可应用于更大规模的生物反应器、珠子或支架。