NDLI: Isolation of murine lymph node stromal cells.

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Year: 2014 Issue: 89

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Isolation of murine lymph node stromal cells.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Broggi, Maria A. S. Schmaler, Mathias Lagarde, Nadège Rossi, Simona W.
Description	Author Affiliation: Broggi MA ( Department of Biomedicine, Immunoregulation, University of Basel and University Hospital Basel.); Schmaler M ( Department of Biomedicine, Immunoregulation, University of Basel and University Hospital Basel.); Lagarde N ( Department of Biomedicine, Immunoregulation, University of Basel and University Hospital Basel.); Rossi SW ( Department of Biomedicine, Immunoregulation, University of Basel and University Hospital Basel)
Abstract	Secondary lymphoid organs including lymph nodes are composed of stromal cells that provide a structural environment for homeostasis, activation and differentiation of lymphocytes. Various stromal cell subsets have been identified by the expression of the adhesion molecule CD31 and glycoprotein podoplanin (gp38), T zone reticular cells or fibroblastic reticular cells, lymphatic endothelial cells, blood endothelial cells and FRC-like pericytes within the double negative cell population. For all populations different functions are described including, separation and lining of different compartments, attraction of and interaction with different cell types, filtration of the draining fluidics and contraction of the lymphatic vessels. In the last years, different groups have described an additional role of stromal cells in orchestrating and regulating cytotoxic T cell responses potentially dangerous for the host. Lymph nodes are complex structures with many different cell types and therefore require a appropriate procedure for isolation of the desired cell populations. Currently, protocols for the isolation of lymph node stromal cells rely on enzymatic digestion with varying incubation times; however, stromal cells and their surface molecules are sensitive to these enzymes, which results in loss of surface marker expression and cell death. Here a short enzymatic digestion protocol combined with automated mechanical disruption to obtain viable single cells suspension of lymph node stromal cells maintaining their surface molecule expression is proposed.
File Format	HTM / HTML
e-ISSN	1940087X
DOI	10.3791/51803
Journal	Journal of Visualized Experiments
Issue Number	90
Language	English
Publisher	MyJove Corp.
Publisher Date	2014-08-19
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Physical Sciences Discipline Life Sciences Discipline Medicine Cytological Techniques Lymph Nodes Cytology Stromal Cells Animals Cell Separation Mice Research Support, Non-u.s. Gov't Video-audio Media
Content Type	Text
Resource Type	Article

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