Ragelle Héloïse, Naba Alexandra, Larson Benjamin L, Zhou Fangheng, Prijić Miralem, Whittaker Charles A, Del Rosario Amanda, Langer Robert, Hynes Richard O, Anderson Daniel G
David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02142, USA.
David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02142, USA; Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA; Institute for Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
Biomaterials. 2017 Jun;128:147-159. doi: 10.1016/j.biomaterials.2017.03.008. Epub 2017 Mar 7.
In the stem-cell niche, the extracellular matrix (ECM) serves as a structural support that additionally provides stem cells with signals that contribute to the regulation of stem-cell function, via reciprocal interactions between cells and components of the ECM. Recently, cell-derived ECMs have emerged as in vitro cell culture substrates to better recapitulate the native stem-cell microenvironment outside the body. Significant changes in cell number, morphology and function have been observed when mesenchymal stem cells (MSC) were cultured on ECM substrates as compared to standard tissue-culture polystyrene (TCPS). As select ECM components are known to regulate specific stem-cell functions, a robust characterization of cell-derived ECM proteomic composition is critical to better comprehend the role of the ECM in directing cellular processes. Here, we characterized and compared the protein composition of ECM produced in vitro by bone marrow-derived MSC, adipose-derived MSC and neonatal fibroblasts from different donors, employing quantitative proteomic methods. Each cell-derived ECM displayed a specific and unique matrisome signature, yet they all shared a common set of proteins. We evaluated the biological response of cells cultured on the different matrices and compared them to cells on standard TCPS. The matrices lead to differential survival and gene-expression profiles among the cell types and as compared to TCPS, indicating that the cell-derived ECMs influence each cell type in a different manner. This general approach to understanding the protein composition of different tissue-specific and cell-derived ECM will inform the rational design of defined systems and biomaterials that recapitulate critical ECM signals for stem-cell culture and tissue engineering.
在干细胞生态位中，细胞外基质（ECM）作为一种结构支撑，通过细胞与ECM成分之间的相互作用，额外为干细胞提供有助于调节干细胞功能的信号。最近，细胞衍生的ECM已成为体外细胞培养底物，以更好地模拟体外天然干细胞微环境。与标准组织培养聚苯乙烯（TCPS）相比，当间充质干细胞（MSC）在ECM底物上培养时，观察到细胞数量、形态和功能发生了显著变化。由于已知某些ECM成分可调节特定的干细胞功能，因此对细胞衍生的ECM蛋白质组组成进行全面表征对于更好地理解ECM在指导细胞过程中的作用至关重要。在这里，我们采用定量蛋白质组学方法，对来自不同供体的骨髓来源的MSC、脂肪来源的MSC和新生儿成纤维细胞在体外产生的ECM的蛋白质组成进行了表征和比较。每种细胞衍生的ECM都显示出特定且独特的基质组特征，但它们都共享一组共同的蛋白质。我们评估了在不同基质上培养的细胞的生物学反应，并将其与在标准TCPS上培养的细胞进行了比较。这些基质导致不同细胞类型之间以及与TCPS相比具有不同的存活和基因表达谱，表明细胞衍生的ECM以不同方式影响每种细胞类型。这种理解不同组织特异性和细胞衍生的ECM蛋白质组成的一般方法将为合理设计能够重现干细胞培养和组织工程关键ECM信号的特定系统和生物材料提供信息。
