Isolation of Uterine Innate Lymphoid Cells for Analysis by Flow Cytometry.
Accepted version
Peer-reviewed
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Repository DOI
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Authors
Depierreux, Delphine M
Seshadri, Emily
Shmeleva, Evgeniya V
Kieckbusch, Jens
Hawkes, Delia A
Abstract
Described here is a simple method to isolate and phenotype mouse group 1 uterine innate lymphoid cells (g1 uILCs) from individual pregnant uterus by flow cytometry. The protocol describes how to set up time mating to obtain multiple synchronous dams, the mechanical and enzymatic digestion of the pregnant uterus, the staining of single-cell suspensions, and a FACS strategy to phenotype and discriminate g1 uILCs. Although this method inevitably loses the spatial information of cellular distribution within the tissue, the protocol has been successfully applied to determine uILC heterogeneity, their response to maternal and foetal factors affecting pregnancy, their gene expression profile, and their functions.
Description
Keywords
Animals, Female, Flow Cytometry, Immunity, Innate, Lymphocytes, Mice, Phenotype, Pregnancy, Uterus
Journal Title
J Vis Exp
Conference Name
Journal ISSN
1940-087X
1940-087X
1940-087X
Volume Title
Publisher
MyJove Corporation
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Rights
All rights reserved
Sponsorship
Medical Research Council (MR/P001092/1)
Wellcome Trust (200841/Z/16/Z)
Wellcome Trust (200841/Z/16/Z)
Our lab is supported by a Wellcome Investigator Award (200841/Z/16/Z), and an MRC project grant (MR/P001092/1)