Department of Stem Cell and Regenerative Medicine, Yenepoya Research Center, Yenepoya University, University Road, Mangalore 575018, Karnataka, India.
Methods. 2017 Dec 1;131:147-156. doi: 10.1016/j.ymeth.2017.06.035. Epub 2017 Jul 4.
Mesenchymal stem cells (MSCs) constitute the diverse progenitor populations in almost every tissue and are of immense importance in the field of regenerative medicine. CD34 is a cell surface glycoprotein identified first as a marker for the MSCs of hematopoietic origin. CD34 is now known to be expressed in cells of diverse lineages (tissues of non-hematopoietic origin) such as ectoderm, mesoderm and endoderm and is considered as a general marker for progenitor cells. Here, we present detailed protocols to obtain pure populations of MSCs from three diverse lineages such as skeletal muscle, skin, and liver from mouse tissues. We also present here the protocol for systems biology approach (proteomic analysis) of these purified cells. This proteomic approach can elucidate key signalling pathways and proteins utilized by these CD34 positive cells in undifferentiated and differentiated conditions. Furthermore in-depth proteomic analysis can also identify the altered proteome which is responsible for their function during non-clinical and clinical conditions.
间充质干细胞（MSCs）构成了几乎每种组织中多样化的祖细胞群体，在再生医学领域具有重要意义。CD34 是一种细胞表面糖蛋白，最初被确定为造血来源的 MSCs 的标志物。现在已知 CD34 表达于不同谱系的细胞（非造血来源的组织）中，如外胚层、中胚层和内胚层，并被认为是祖细胞的通用标志物。在这里，我们提供了从三种不同谱系（如骨骼肌、皮肤和肝脏）的小鼠组织中获得纯 MSCs 群体的详细方案。我们还介绍了针对这些纯化细胞的系统生物学方法（蛋白质组学分析）的方案。这种蛋白质组学方法可以阐明这些 CD34 阳性细胞在未分化和分化条件下利用的关键信号通路和蛋白质。此外，深入的蛋白质组学分析还可以识别负责其在非临床和临床条件下功能的改变蛋白质组。
