Lalwani Gaurav, D'agati Michael, Gopalan Anu, Patel Sunny C, Talukdar Yahfi, Sitharaman Balaji
Department of Biomedical Engineering, Stony Brook University, Stony Brook, New York, 11794-5281.
J Biomed Mater Res A. 2017 Jul;105(7):1927-1939. doi: 10.1002/jbm.a.36062. Epub 2017 Apr 3.
Expansion of mesenchymal stem cells (MSCs) and maintenance of their self-renewal capacity in vitro requires specialized robust cell culture systems. Conventional approaches using animal-derived or artificial matrices and a cocktail of growth factors have limitations such as consistency, scalability, pathogenicity, and loss of MSC phenotype. Herein, we report the use of all-carbon 3-D single- and multiwalled carbon nanotube scaffolds (SWCNTs and MWCNTs) as artificial matrices for long-term maintenance and expansion of human MSCs. Three-dimensional SWCNT and MWCNT scaffolds were fabricated using a novel radical initiated thermal cross-linking method that covalently cross-links CNTs to form 3-D macroporous all-carbon architectures. Adipose-derived human MSCs showed good cell viability, attachment, proliferation, and infiltration in MWCNT and SWCNT scaffolds comparable to poly(lactic-co-glycolic) acid (PLGA) scaffolds (baseline control). ADSCs retained stem cell phenotype after 30 days and satisfied the International Society for Cellular Therapy's (ISCT) minimal criteria for MSCs. Post expansion, (1) ADSCs showed in vitro adherence to tissue culture polystyrene (TCPS); (2) MSC surface antigen expression [CD14(-), CD19(-), CD34(-), CD45(-), CD73(+), CD90(+), CD105(+)]; and (3) trilineage differentiation into osteoblasts, adipocytes, and chondrocytes. Results show that cross-linked 3-D MWCNTs and SWCNTs scaffolds are suitable for ex vivo expansion and maintenance of MSCs for therapeutic applications. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1927-1939, 2017.
间充质干细胞（MSCs）的体外扩增及其自我更新能力的维持需要专门的强大细胞培养系统。使用动物源或人工基质以及生长因子混合物的传统方法存在局限性，如一致性、可扩展性、致病性以及MSCs表型丧失等问题。在此，我们报告使用全碳三维单壁和多壁碳纳米管支架（SWCNTs和MWCNTs）作为人工基质用于人MSCs的长期维持和扩增。三维SWCNT和MWCNT支架采用一种新型自由基引发热交联方法制备，该方法通过共价交联碳纳米管形成三维大孔全碳结构。脂肪来源的人MSCs在MWCNT和SWCNT支架中表现出良好的细胞活力、附着、增殖和浸润能力，与聚乳酸-乙醇酸共聚物（PLGA）支架（基线对照）相当。脂肪来源的干细胞（ADSCs）在30天后保留了干细胞表型，并满足国际细胞治疗协会（ISCT）对MSCs的最低标准。扩增后，（1）ADSCs在体外可附着于组织培养聚苯乙烯（TCPS）；（2）MSC表面抗原表达[CD14(-)、CD19(-)、CD34(-)、CD45(-)、CD73(+)、CD90(+)、CD105(+)]；（3）可向成骨细胞、脂肪细胞和软骨细胞进行三系分化。结果表明，交联的三维MWCNTs和SWCNTs支架适用于MSCs的体外扩增和维持，以用于治疗应用。© 2016威利期刊公司。《生物医学材料研究杂志》A部分：105A：1927 - 1939，2017年。
