title: [Isolation, primary culture and characterization of mouse glomerular mesangial cells].
abstract: OBJECTIVE
  To establish a method for the isolation, primary culture and characterization of mouse mesangial cells (MCs).
  
  
  METHODS
  We used two-layer micropore filter device to isolate glomeruli, and the MCs were cultured with eugenic selection method. Cell structures of MCs were observed by inverted phase contrast microscope, HE staining, scanning electron microscope and transmission electron microscope. Expressions of α-smooth muscle actin (α-SMA and nephrin were investigated by immunofluorescent cytochemistry. We used angiotensin II to stimulate MCs and observed the biological characteristics of the cells. The growth curve of MCs was examined by CCK-8 assay.
  
  
  RESULTS
  After 20-30 days, the primary cultured MCs gradually covered the bottom of the dish. The mouse MCs usually attached to the surface 6-10 hours after passage, followed by the exponential phase in 2-3 days and the platform period in 3-5 days. Immunofluorescent staining showed that the expression of α-SMA was positive and nephrin was negative. MCs contraction was observed after the cells were stimulated by angiotensin II for 15 minutes.
  
  
  CONCLUSIONS
  A method for the isolation, culture and characterization of mouse mesangial cells has been successfully established.
authors: Department of Nephrology, Affiliated Hospital, Nantong University, Nantong 226001, China.
authors: Yingjie
authors: Y
authors: Cao
authors: Naifeng
authors: N
authors: Guo
authors: Ying
authors: Y
authors: Lv
authors: Hui
authors: H
authors: Shi
authors: Xiaolan
authors: X
authors: Chen
mesh_terms: D000804:Angiotensin II / Q000494:pharmacology; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation / Q000187:drug effects; D002469:Cell Separation / Q000379:methods*; D050527:Mesangial Cells / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D051379:Mice; D008810:Mice, Inbred C57BL; D008854:Microscopy, Electron
chemical_list: D000804:Angiotensin II