School of Ophthalmology and Optometry, The Eye Hospital of Wenzhou Medical University, No. 270, Xueyuan Road, Wenzhou, Zhejiang 325027, China.
Department of Chemical and Materials Engineering, National Central University, No. 300, Jhongda Rd, Jhongli, Taoyuan 32001, Taiwan.
J Mater Chem B. 2020 Jun 24;8(24):5204-5214. doi: 10.1039/d0tb00947d.
Human adipose-derived stem cells (hASCs) cultured for 5 passages were filtered through nylon (NY) mesh filter membranes coated with and without extracellular matrix proteins to obtain the permeation solution. Subsequently, the culture media were filtered via the membranes to obtain the recovery solution. Then, the membranes were cultured in cell culture medium to obtain the migrated cells from the membranes. The hASCs in the permeation solution, through any type of NY mesh filter membrane having 11 and 20 μm pore sizes, had lower osteogenic differentiation ability than conventional hASCs cultured on tissue culture polystyrene (TCP) dishes for passage 5, whereas the hASCs purified by the membrane migration method through NY mesh filter membranes coated with recombinant vitronectin, which have 11 and 20 μm pore sizes, showed a higher proliferation speed as well as higher osteogenic differentiation potential than the conventional hASCs cultured on TCP dishes for passage 5. The membrane filtration and migration methods would be useful for cell sorting for specific cells, such as hASCs with high proliferation and high osteogenic differentiation ability, which do not need antibody binding or genetic modification of the cells for the specific isolation of the cells.
培养 5 代的人脂肪干细胞(hASCs)通过尼龙(NY)滤网过滤,滤网涂有或不涂细胞外基质蛋白,以获得渗透溶液。随后,通过膜过滤培养基,获得回收溶液。然后,将膜在细胞培养基中培养,从膜中获得迁移细胞。与在组织培养聚苯乙烯(TCP)板上培养至第 5 代的常规 hASCs 相比,通过具有 11 和 20μm 孔径的任何类型 NY 滤网过滤的渗透溶液中的 hASCs 具有较低的成骨分化能力,而通过涂有重组 vitronectin 的 NY 滤网过滤膜进行膜迁移法纯化的 hASCs 具有较高的增殖速度和更高的成骨分化潜能比在 TCP 板上培养至第 5 代的常规 hASCs。膜过滤和迁移方法可用于特定细胞的细胞分选,例如具有高增殖和高成骨分化能力的 hASCs,而不需要为了特定的细胞分离而对细胞进行抗体结合或基因修饰。