Epshtein Alona, Sakhneny Lina, Landsman Limor
Cell and Developmental Biology, Sackler Faculty of Medicine, Tel Aviv University.
Cell and Developmental Biology, Sackler Faculty of Medicine, Tel Aviv University;
J Vis Exp. 2017 Jan 28(119):55344. doi: 10.3791/55344.
The pancreas is comprised of epithelial cells that are required for food digestion and blood glucose regulation. Cells of the pancreas microenvironment, including endothelial, neuronal, and mesenchymal cells were shown to regulate cell differentiation and proliferation in the embryonic pancreas. In the adult, the function and mass of insulin-producing cells were shown to depend on cells in their microenvironment, including pericyte, immune, endothelial, and neuronal cells. Lastly, changes in the pancreas microenvironment were shown to regulate pancreas tumorigenesis. However, the cues underlying these processes are not fully defined. Therefore, characterizing the different cell types that comprise the pancreas microenvironment and profiling their gene expression are crucial to delineate the tissue development and function under normal and diseased states. Here, we describe a method that allows for the isolation of mesenchymal cells from the pancreas of embryonic, neonatal, and adult mice. This method utilizes the enzymatic digestion of mouse pancreatic tissue and the subsequent fluorescence-activated cell sorting (FACS) or flow-cytometric analysis of labeled cells. Cells can be labeled by either immunostaining for surface markers or by the expression of fluorescent proteins. Cell isolation can facilitate the characterization of genes and proteins expressed in cells of the pancreas mesenchyme. This protocol was successful in isolating and culturing highly enriched mesenchymal cell populations from the embryonic, neonatal, and adult mouse pancreas.
胰腺由食物消化和血糖调节所需的上皮细胞组成。胰腺微环境中的细胞，包括内皮细胞、神经元细胞和间充质细胞，已被证明在胚胎胰腺中调节细胞分化和增殖。在成体中，产生胰岛素的细胞的功能和质量已被证明取决于其微环境中的细胞，包括周细胞、免疫细胞、内皮细胞和神经元细胞。最后，胰腺微环境的变化已被证明可调节胰腺肿瘤发生。然而，这些过程背后的线索尚未完全明确。因此，表征构成胰腺微环境的不同细胞类型并分析它们的基因表达对于描绘正常和疾病状态下的组织发育和功能至关重要。在这里，我们描述了一种从胚胎、新生和成年小鼠的胰腺中分离间充质细胞的方法。该方法利用小鼠胰腺组织的酶消化以及随后对标记细胞的荧光激活细胞分选（FACS）或流式细胞术分析。细胞可以通过表面标志物的免疫染色或荧光蛋白的表达进行标记。细胞分离有助于表征胰腺间充质细胞中表达的基因和蛋白质。该方案成功地从胚胎、新生和成年小鼠胰腺中分离并培养了高度富集的间充质细胞群体。
