title: Reliable and High Efficiency Extraction of Kidney Immune Cells.
abstract: Immune system activation occurs in multiple kidney diseases and pathophysiological processes. The immune system consists of both adaptive and innate components and multiple cell types. Sometimes, the cell type of interest is present in very low numbers among the large numbers of total cells isolated from the kidney. Hence, reliable and efficient isolation of kidney mononuclear cell populations is important in order to study the immunological problems associated with kidney diseases. Traditionally, tissue isolation of kidney mononuclear cells have been performed via enzymatic digestions using different varieties and strengths of collagenases/DNAses yielding varying numbers of viable immune cells. Recently, with the development of the mechanical tissue disruptors for single cell isolation, the collagenase digestion step is avoided and replaced by a simple mechanical disruption of the kidneys after extraction from the mouse. Herein, we demonstrate a simple yet efficient method for the isolation of kidney mononuclear cells for every day immune cell extractions. We further demonstrate an example of subset analysis of immune cells in the kidney. Importantly, this technique can be adapted to other soft and non-fibrous tissues such as the liver and brain.
authors: Division of Nephrology, Department of Medicine, University of Missouri-Columbia and Harry S Truman Memorial Veteran's Hospital; nistalar@health.missouri.edu.
authors: Ravi
authors: R
authors: Nistala
authors: Division of Biomedical Sciences, Department of Medicine, University of Missouri-Columbia and Harry S Truman Memorial Veteran's Hospital.
authors: Alex
authors: A
authors: Meuth
authors: Division of Endocrinology, Department of Medicine, University of Missouri-Columbia and Harry S Truman Memorial Veteran's Hospital.
authors: Cassandra
authors: C
authors: Smith
authors: Division of Endocrinology, Department of Medicine, University of Missouri-Columbia and Harry S Truman Memorial Veteran's Hospital.
authors: Aroor
authors: A
authors: Annayya
mesh_terms: D000818:Animals; D015415:Biomarkers; D002469:Cell Separation / Q000379:methods*; D002499:Centrifugation, Density Gradient; D005434:Flow Cytometry; D007107:Immune System / Q000166:cytology*; D007150:Immunohistochemistry; D007668:Kidney / Q000166:cytology* / Q000276:immunology*; D008264:Macrophages / Q000166:cytology* / Q000276:immunology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D015203:Reproducibility of Results; D016176:T-Lymphocyte Subsets / Q000166:cytology* / Q000276:immunology
chemical_list: D015415:Biomarkers