Isolation of Uterine Innate Lymphoid Cells for Analysis by Flow Cytometry. 2021

Delphine M Depierreux, and Emily Seshadri, and Evgeniya V Shmeleva, and Jens Kieckbusch, and Delia A Hawkes, and Francesco Colucci
Department of Obstetrics and Gynaecology, National Institute for Health Research Cambridge Biomedical Research Centre, University of Cambridge School of Clinical Medicine; Centre for Trophoblast Research, University of Cambridge.

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.

UI MeSH Term Description Entries
D007113 Immunity, Innate The capacity of a normal organism to remain unaffected by microorganisms and their toxins. It results from the presence of naturally occurring ANTI-INFECTIVE AGENTS, constitutional factors such as BODY TEMPERATURE and immediate acting immune cells such as NATURAL KILLER CELLS. Immunity, Native,Immunity, Natural,Immunity, Non-Specific,Resistance, Natural,Innate Immune Response,Innate Immunity,Immune Response, Innate,Immune Responses, Innate,Immunity, Non Specific,Innate Immune Responses,Native Immunity,Natural Immunity,Natural Resistance,Non-Specific Immunity
D008214 Lymphocytes White blood cells formed in the body's lymphoid tissue. The nucleus is round or ovoid with coarse, irregularly clumped chromatin while the cytoplasm is typically pale blue with azurophilic (if any) granules. Most lymphocytes can be classified as either T or B (with subpopulations of each), or NATURAL KILLER CELLS. Lymphoid Cells,Cell, Lymphoid,Cells, Lymphoid,Lymphocyte,Lymphoid Cell
D010641 Phenotype The outward appearance of the individual. It is the product of interactions between genes, and between the GENOTYPE and the environment. Phenotypes
D011247 Pregnancy The status during which female mammals carry their developing young (EMBRYOS or FETUSES) in utero before birth, beginning from FERTILIZATION to BIRTH. Gestation,Pregnancies
D005260 Female Females
D005434 Flow Cytometry Technique using an instrument system for making, processing, and displaying one or more measurements on individual cells obtained from a cell suspension. Cells are usually stained with one or more fluorescent dyes specific to cell components of interest, e.g., DNA, and fluorescence of each cell is measured as it rapidly transverses the excitation beam (laser or mercury arc lamp). Fluorescence provides a quantitative measure of various biochemical and biophysical properties of the cell, as well as a basis for cell sorting. Other measurable optical parameters include light absorption and light scattering, the latter being applicable to the measurement of cell size, shape, density, granularity, and stain uptake. Cytofluorometry, Flow,Cytometry, Flow,Flow Microfluorimetry,Fluorescence-Activated Cell Sorting,Microfluorometry, Flow,Cell Sorting, Fluorescence-Activated,Cell Sortings, Fluorescence-Activated,Cytofluorometries, Flow,Cytometries, Flow,Flow Cytofluorometries,Flow Cytofluorometry,Flow Cytometries,Flow Microfluorometries,Flow Microfluorometry,Fluorescence Activated Cell Sorting,Fluorescence-Activated Cell Sortings,Microfluorimetry, Flow,Microfluorometries, Flow,Sorting, Fluorescence-Activated Cell,Sortings, Fluorescence-Activated Cell
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D014599 Uterus The hollow thick-walled muscular organ in the female PELVIS. It consists of the fundus which is the site of EMBRYO IMPLANTATION and FETAL DEVELOPMENT. Beyond the isthmus at the perineal end of fundus, is CERVIX UTERI (the neck) opening into VAGINA. Beyond the isthmi at the upper abdominal end of fundus, are the FALLOPIAN TUBES. Fundus Uteri,Uteri,Uterine Cornua,Uterine Fundus,Uterus Cornua,Womb,Cornua, Uterine,Fundus Uterus,Fundus, Uterine,Uteri, Fundus,Wombs
D051379 Mice The common name for the genus Mus. Mice, House,Mus,Mus musculus,Mice, Laboratory,Mouse,Mouse, House,Mouse, Laboratory,Mouse, Swiss,Mus domesticus,Mus musculus domesticus,Swiss Mice,House Mice,House Mouse,Laboratory Mice,Laboratory Mouse,Mice, Swiss,Swiss Mouse,domesticus, Mus musculus

Related Publications

Delphine M Depierreux, and Emily Seshadri, and Evgeniya V Shmeleva, and Jens Kieckbusch, and Delia A Hawkes, and Francesco Colucci
January 2017, Methods in molecular biology (Clifton, N.J.),
Delphine M Depierreux, and Emily Seshadri, and Evgeniya V Shmeleva, and Jens Kieckbusch, and Delia A Hawkes, and Francesco Colucci
January 2014, PloS one,
Delphine M Depierreux, and Emily Seshadri, and Evgeniya V Shmeleva, and Jens Kieckbusch, and Delia A Hawkes, and Francesco Colucci
March 2023, STAR protocols,
Delphine M Depierreux, and Emily Seshadri, and Evgeniya V Shmeleva, and Jens Kieckbusch, and Delia A Hawkes, and Francesco Colucci
November 2014, Current protocols in immunology,
Delphine M Depierreux, and Emily Seshadri, and Evgeniya V Shmeleva, and Jens Kieckbusch, and Delia A Hawkes, and Francesco Colucci
October 1992, Kidney international,
Delphine M Depierreux, and Emily Seshadri, and Evgeniya V Shmeleva, and Jens Kieckbusch, and Delia A Hawkes, and Francesco Colucci
February 2023, Cytometry. Part A : the journal of the International Society for Analytical Cytology,
Delphine M Depierreux, and Emily Seshadri, and Evgeniya V Shmeleva, and Jens Kieckbusch, and Delia A Hawkes, and Francesco Colucci
August 2018, Current protocols in immunology,
Delphine M Depierreux, and Emily Seshadri, and Evgeniya V Shmeleva, and Jens Kieckbusch, and Delia A Hawkes, and Francesco Colucci
January 2023, Frontiers in immunology,
Delphine M Depierreux, and Emily Seshadri, and Evgeniya V Shmeleva, and Jens Kieckbusch, and Delia A Hawkes, and Francesco Colucci
January 2014, Methods in enzymology,
Delphine M Depierreux, and Emily Seshadri, and Evgeniya V Shmeleva, and Jens Kieckbusch, and Delia A Hawkes, and Francesco Colucci
January 2006, Nature protocols,
Copied contents to your clipboard!