Isolation of functional dendritic cells from murine kidneys for immunological characterization

Nephrology (Carlton). 2012 May;17(4):364-71. doi: 10.1111/j.1440-1797.2012.01581.x.

Abstract

Aim: The kidney is a complex organ, requiring the contributions of multiple cell types to perform its various functions. Within this system the dendritic cell has been demonstrated to play a key role in maintaining the immunological balance of the kidney. In this methods paper we aim to identify the best method for isolating murine renal dendritic cells.

Methods: The efficiency of isolating dendritic cells from enzymatically digested renal parenchyma by density centrifugation, MACS and FACS was compared.

Results: Density centrifugation enriched dendritic cells by only approximately two fold. However, MACS and FACS resulted in a much higher purity (80% versus 95% respectively).

Conclusions: Although FACS gave the highest purity, MACS is the optimal method for isolating dendritic cells given cost and time factors. Isolation of a homogeneous population of renal dendritic cells will enable the molecular and functional dissection of these cells in both homeostasis and disease models.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biomarkers / analysis
  • Cell Count
  • Cell Separation* / methods
  • Centrifugation, Density Gradient
  • Dendritic Cells / immunology*
  • Flow Cytometry
  • Immunomagnetic Separation
  • Kidney / cytology
  • Kidney / immunology*
  • Mice
  • Mice, Inbred C57BL

Substances

  • Biomarkers