Department of Orthopedics and Rehabilitation, Laboratory of Musculoskeletal Biology and Regenerative Medicine, University of Wisconsin-Madison, 1111 Highland Avenue, WIMR 5051, Madison, WI, 53705-2275, USA.
Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, USA.
Stem Cell Rev Rep. 2016 Oct;12(5):560-572. doi: 10.1007/s12015-016-9671-7.
Stem cell fate is largely determined by the microenvironment called niche. The extracellular matrix (ECM), as a key component in the niche, is responsible for maintaining structural stability and regulating cell proliferation, differentiation, migration and other cellular activities. Each tissue has a unique ECM composition for its needs. Here we investigated the effect of a bioengineered human dermal fibroblast-derived ECM (hECM) on the regulation of human mesenchymal stem cell (hMSC) proliferation and multilineage differentiation. Human MSCs were maintained on hECM for two passages followed by the analysis of mRNA expression levels of potency- and lineage-specific markers to determine the capacity of MSC stemness and differentiation, respectively. Mesenchymal stem cells pre-cultured with or without hECM were then induced and analyzed for osteogenesis, adipogenesis and chondrogenesis. Our results showed that compared to MSCs maintained on control culture plates without hECM coating, cells on hECM-coated plates proliferated more rapidly with a higher percentage of cells in S phase of the cell cycle, resulting in an increase in the CD90/CD105/CD73/CD45 subpopulation. In addition, hECM downregulated osteogenesis and adipogenesis of hMSCs but significantly upregulated chondrogenesis with increased production of collagen type 2. In sum, our findings suggest that hECM may be used to culture hMSCs for the application of cartilage tissue engineering.
干细胞命运在很大程度上取决于称为龛的微环境。细胞外基质 (ECM) 作为龛的一个关键组成部分,负责维持结构稳定性并调节细胞增殖、分化、迁移和其他细胞活动。每种组织都有其独特的 ECM 组成以满足其需求。在这里,我们研究了生物工程人真皮成纤维细胞衍生的 ECM(hECM)对人间充质干细胞(hMSC)增殖和多能分化的调节作用。人 MSC 在 hECM 上维持两个传代,然后分析效力和谱系特异性标志物的 mRNA 表达水平,以分别确定 MSC 干性和分化的能力。然后用或不用 hECM 预培养间充质干细胞,并诱导其进行成骨、成脂和成软骨分化分析。我们的结果表明,与在没有 hECM 涂层的对照培养板上维持的 MSC 相比,在 hECM 涂层板上培养的细胞增殖更快,细胞周期 S 期的细胞比例更高,导致 CD90/CD105/CD73/CD45 亚群增加。此外,hECM 下调 hMSCs 的成骨和成脂分化,但显著上调软骨分化,增加 II 型胶原蛋白的产生。总之,我们的研究结果表明,hECM 可用于培养 hMSC 以应用于软骨组织工程。