Cooperative Research Centre for Cell Therapy Manufacturing , North Terrace, Adelaide, South Australia 5000, Australia.
Future Industries Institute, University of South Australia , Mawson Lakes, South Australia 5095, Australia.
ACS Appl Mater Interfaces. 2017 Feb 1;9(4):3445-3454. doi: 10.1021/acsami.6b14725. Epub 2017 Jan 19.
Culture surfaces that substantially reduce the degree of cell manipulation in the delivery of cell sheets to patients are described. These surfaces support the attachment, culture, and delivery of multipotent adult progenitor cells (MAPC). It was essential that the processes of attachment/detachment to the surface did not affect cell phenotype nor the function of the cultured cells. Both acid-based and amine-based surface coatings were generated from acrylic acid, propanoic acid, diaminopropane, and heptylamine precursors, respectively. While both functional groups supported cell attachment/detachment, amine coated surfaces gave optimal performance. X-ray photoelectron spectroscopy (XPS) showed that at a primary amine to carbon surface ratio of between 0.01 and 0.02, greater than 90% of attached cells were effectively transferred to a model wound bed. A dependence on primary amine concentration has not previously been reported. After 48 h of culture on the optimized amine surface, PCR, functional, and viability assays showed that MAPC retained their stem cell phenotype, full metabolic activity, and biological function. Consequently, in a proof of concept experiment, it was shown that this amine surface when coated onto a surgical dressing provides an effective and simple technology for the delivery of MAPC to murine dorsal excisional wounds, with MAPC delivery verified histologically. By optimizing for cell delivery using a combination of in vitro and in vivo techniques, we developed an effective surface for the delivery of MAPC in a clinically relevant format.
描述了可大大减少细胞片在递送至患者过程中细胞操作程度的培养表面。这些表面支持多能成体祖细胞(MAPC)的附着、培养和递送。至关重要的是,附着/从表面脱离的过程不会影响细胞表型或培养细胞的功能。酸基和胺基表面涂层分别由丙烯酸、丙酸、二氨基丙烷和庚胺前体制成。虽然两种官能团都支持细胞附着/脱离,但胺涂层表面的性能最佳。X 射线光电子能谱(XPS)表明,在伯胺与碳表面比率为 0.01 到 0.02 之间时,超过 90%的附着细胞可有效地转移到模型创面床。此前尚未报道过对伯胺浓度的依赖性。在优化的胺表面上培养 48 小时后,PCR、功能和活力检测表明 MAPC 保留了其干细胞表型、完全的代谢活性和生物学功能。因此,在概念验证实验中,表明当这种胺表面涂覆到手术敷料上时,它为 MAPC 递送至鼠背切除伤口提供了一种有效且简单的技术,并且可以通过组织学验证 MAPC 的递送。通过结合体外和体内技术优化细胞递送,我们开发了一种有效的表面,以临床相关的形式递送 MAPC。