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Process and Analysis of Kidney Infiltrates by Flow Cytometry from Murine Lupus Nephritis

从狼疮性肾炎小鼠模型中分离肾脏炎症细胞及流式细胞术分析

Ramalingam Bethunaickan Ramalingam Bethunaickan
AD Anne Davidson

发布: 2012年05月05日第2卷第9期 DOI: 10.21769/BioProtoc.167 浏览次数: 17934

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Abstract

Methods for the isolation and characterization of mononuclear phagocytes from the kidneys of mice with SLE are essential to understand the patho-physiology of the disease. Activation of these cells is associated with the onset of clinical disease in mice and infiltration with these cells is associated with poor prognosis in humans.An analysis of the function of these cells should lead to a better understanding of the inflammatory processes that lead to renal impairment in SLE and other renal inflammatory diseases.

Keywords: SLE nephritis (SLE肾炎) search Kidney infiltrates (肾infiltrates) search Macrophages from kidney (肾macrophages from) search Collagen digestion (胶原消化) search Inflammation (炎症) search

Materials and Reagents

  1. Fetal bovine serum (FBS)
  2. Sterile PBS (Life Technologies, Invitrogen™, catalog number: 20012-027 )
  3. 0.17M Ammonium chloride
  4. Collagenase Type I (CLS I) (Worthington, catalog number: 4197 , specific activity 230 U mg-1)
  5. DMEM, High glucose (Life Technologies, Gibco®, catalog number: 10313 )
  6. Paraformaldehyde (Tousimis, catalog number: 1108A )
  7. BSA (Fraction V) (Sigma-Aldrich, catalog number: A7030 )
  8. Fc block (CD16/CD32)
  9. FACS staining buffer (see Recipes)

Equipment

  1. BD LSRII or similar flow cytometer
  2. Bench-top refrigerated centrifuge
  3. BD cell strainer (40 nm) (BD Biosciences, Falcon®, catalog number: 352340 )
  4. 30 ml syringe (BD Biosciences, Falcon®, catalog number: 309661 )
  5. Microscopes
  6. 21G Needles (BD Biosciences, Falcon®, catalog number: 305165 )
  7. 26G needles (BD Biosciences, Falcon®, catalog number: 305111 )
  8. V bottom 96 well Assay plate (Corning, Costar® , catalog number: 3897 )
  9. Glass slides Frosted (Thermo Fisher Scientific, catalog number: 12-550-11 )

Procedure

English
中文翻译

文章信息

版权信息

© 2012 The Authors; exclusive licensee Bio-protocol LLC.

如何引用

Bethunaickan, R. and Davidson, A. (2012). Process and Analysis of Kidney Infiltrates by Flow Cytometry from Murine Lupus Nephritis. Bio-protocol 2(9): e167. DOI: 10.21769/BioProtoc.167.

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分类

免疫学 > 免疫细胞功能 > 综合

细胞生物学 > 基于细胞的分析方法 > 流式细胞术

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