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J Vis Exp. 2017 Jan 28;(119). doi: 10.3791/55344.

Isolating and Analyzing Cells of the Pancreas Mesenchyme by Flow Cytometry.

Author information

1
Cell and Developmental Biology, Sackler Faculty of Medicine, Tel Aviv University.
2
Cell and Developmental Biology, Sackler Faculty of Medicine, Tel Aviv University; limorl@post.tau.ac.il.

Abstract

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.

PMID:
28190046
PMCID:
PMC5352303
DOI:
10.3791/55344
[Indexed for MEDLINE]
Free PMC Article

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