Center for Biomaterials, Korea Institute of Science and Technology (KIST), Seoul, 02792, Republic of Korea.
Department of Biotechnology, Korea University, Seoul, 02841, Republic of Korea.
Cell Tissue Res. 2019 May;376(2):233-245. doi: 10.1007/s00441-018-2971-2. Epub 2019 Jan 4.
Large expansion of human mesenchymal stem cells (MSCs) is of great interest for clinical applications. In this study, we examine the feasibility of human fibroblast-derived extracellular matrix (hFDM) as an alternative cell expansion setting. hFDM is obtained from decellularized extracellular matrix (ECM) derived from in vitro cultured human lung fibroblasts. Our study directly compares conventional platforms (tissue culture plastic (TCP), fibronectin (FN)-coated TCP) with hFDM using umbilical cord blood-derived MSCs (UCB-MSCs). Early cell morphology shows a rather rounded shape on TCP but highly elongated morphology on hFDM. Cell proliferation demonstrates that MSCs on hFDM were significantly better compared to the others in both 10 and 2% serum condition. Cell migration assay suggests that cell motility was improved and a cell migration marker CXCR4 was notably up-regulated on hFDM. MSCs differentiation into osteogenic lineage on hFDM was also very effective as examined via gene expression, von Kossa staining and alkaline phosphatase activity. In addition, as the MSCs were expanded on each substrate, transferred to 3D polymer mesh scaffolds and then cultivated for a while, the data found better cell proliferation and more CXCR4 expression with MSCs pre-conditioned on hFDM. Moreover, higher gene expression of stemness and engraftment-related markers was noticed with the hFDM group. Furthermore when UCB-MSCs expanded on TCP or hFDM were injected into emphysema (a lung disease) animal model, the results indicate that MSCs pre-conditioned on hFDM (with 2% serum) retain more advanced therapeutic efficacy on the improvement of emphysema than those on TCP. Current works demonstrate that compared to the conventional platforms, hFDM can be a promising source of cell expansion with a naturally derived biomimetic ECM microenvironment and may find some practical applications in regenerative medicine.
大量扩增人骨髓间充质干细胞（MSCs）对于临床应用具有重要意义。在本研究中，我们研究了人纤维母细胞衍生细胞外基质（hFDM）作为替代细胞扩增体系的可行性。hFDM 是从体外培养的人肺成纤维细胞的去细胞细胞外基质（ECM）中获得的。我们的研究直接比较了传统平台（组织培养塑料（TCP），纤连蛋白（FN）包被的 TCP）与脐带血来源的 MSCs（UCB-MSCs）在 hFDM 上的使用情况。早期细胞形态在 TCP 上呈现出相当圆的形状，但在 hFDM 上呈现出高度伸长的形态。细胞增殖表明，在 10%和 2%血清条件下，hFDM 上的 MSC 增殖明显优于其他平台。细胞迁移实验表明，细胞迁移能力提高，细胞迁移标志物 CXCR4 在 hFDM 上显著上调。通过基因表达、Von Kossa 染色和碱性磷酸酶活性检测，hFDM 上 MSC 向成骨谱系分化也非常有效。此外，当将 MSCs 在每种基质上扩增，转移到 3D 聚合物网格支架上并培养一段时间后，发现 hFDM 预处理的 MSCs 具有更好的细胞增殖和更多的 CXCR4 表达。此外，hFDM 组注意到更高的干性和植入相关标记物的基因表达。此外，当将在 TCP 或 hFDM 上扩增的 UCB-MSCs 注射到肺气肿（一种肺部疾病）动物模型中时，结果表明，在 TCP 上预处理的 hFDM（含 2%血清）的 MSC 保留了更先进的治疗肺气肿的疗效，优于 TCP 上的 MSC。目前的工作表明，与传统平台相比，hFDM 可以成为细胞扩增的有前途的来源，具有天然衍生的仿生细胞外基质微环境，并可能在再生医学中找到一些实际应用。
