Adenoviral-mediated transfer of TGF-β1 but not IGF-1 induces chondrogenic differentiation of human mesenchymal stem cells in pellet cultures
Adenoviral-mediated transfer of TGF-β1 but not IGF-1 induces chondrogenic differentiation of human mesenchymal stem cells in pellet cultures
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DOI：
10.1016/j.exphem.2005.05.010
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发表时间：
2005-08-01
影响因子：
2.6
通讯作者：
Niyibizi, C
中科院分区：
文献类型：
--
作者：
Kawamura, K;Chu, CR;Niyibizi, C
Objective.本研究的目的是探讨应用生长因子基因诱导人骨髓间充质干细胞（MSCs）向软骨细胞分化的潜力。本研究中评估的生长因子基因是转化生长因子1（TGF-β 1）和胰岛素样生长因子1（IGF-1）。用携带TGF-β 1或IGF-1（分别为AdTGF-β 1和AdIGF-1）或两种生长因子基因的组合的腺病毒载体以不同的感染复数（MOI）转导人MSC，然后制成颗粒。还从未转导的细胞制备沉淀，并维持在补充有10 ng/mL TGF-β 1的培养基中。在特定的时间点，进行组织学分析，软骨基质基因表达，免疫荧光，以确定软骨分化的程度。用AdTGF-β 1转导的MSC表现出强大的软骨分化，而由AdIGF-1制成的MSC则没有。AdTGF-P1小球早在小球培养的第3天就表现出聚集蛋白聚糖基因表达，而II型胶原蛋白基因表达在培养的第10天检测到。单独或与TGF-β 1颗粒组合的AdIGF-1在任何时间点均未显示任何II型胶原基因表达。免疫荧光显示TGF-β 1蛋白颗粒中X型胶原分布于整个基质中，而生长因子基因颗粒中X型胶原染色不明显。结果表明，持续给予TGF-β 1可能比间歇给药更有效地抑制终末分化，因此对软骨修复有效。(c)2005年国际实验血液学学会。爱思唯尔公司出版
Objective. The objective of the present study was to investigate the potential of application of growth factor genes to induce chondrogenic differentiation of human-derived mesenchymal stem cells (MSCs). The growth factor genes evaluated in the present study were transforming growth factor 1 (TGF-beta 1) and insulin-like growth factor 1 (IGF-1).Methods. Human MSCs were transduced with the adenoviral vectors carrying either TGF-beta 1 or IGF-1 (AdTGF-beta 1 and AdIGF-1 respectively) or a combination of both growth factor genes at different multiplicities of infection (MOI) and were then made into pellets. Pellets were also made from nontransduced cells and maintained in culture medium supplemented with 10 ng/mL of TGF-beta 1. At specified time points, histological analysis, cartilage matrix gene expression, and immunofluorescence were performed to determine the extent of chundrogenic differentiation.Results. MSCs transduced with the AdTGF-beta 1 demonstrated robust chondrogenic differentiation, while those made from AdIGF-1 did not. AdTGF-Pl pellets demonstrated aggrecan gene expression as early as day 3 of pellet culture, while type II collagen gene expression was detected by day 10 of culture. The AdIGF-1, alone or in combination with TGF-beta 1 pellets, did not show any type II collagen gene expression at any time point. By immunofluoresecence, type X collagen was distributed throughout the matrix in TGF-beta 1 protein pellets while the growth factor gene pellets displayed scant staining.Conclusion. The results suggest that sustained administration of TGF-beta 1 may be more effective in suppressing terminal differentiation than intermittent dosing and thus effective for cartilage repair. (c) 2005 International Society for Experimental Hematology. Published by Elsevier Inc.