title: Numerical measurement of viable and nonviable adipocytes and other cellular components in aspirated fat tissue.
abstract: BACKGROUND
  A reliable method with which to assay viability and number of adipocytes and other cellular components in adipose tissue remains to be established.
  
  
  METHODS
  The authors assessed cell viability and number obtained from 1 g of suctioned adipose tissue and respective layers (the top, middle, and bottom layers) before and after digestion and centrifugation, using cell staining with Hoechst 33342 and propidium iodide and the 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenyl-amino)carbonyl]-2H-tetrazoliumhydroxide (XTT) and glycerol-3-phosphate dehydrogenase assays (n = 10). The correlation between the number of prepared cells (adipocytes, adipose stromal cells, and white blood cells) and the resulting values from the XTT and glycerol-3-phosphate dehydrogenase assays was also examined (n = 5). The cell composition of the stromal vascular fraction isolated from the same adipose tissue was determined by multicolor flow cytometry (n = 5).
  
  
  RESULTS
  Hoechst 33342 and propidium iodide staining allowed distinguishing of viable adipocytes from lipid droplets, dead adipocytes, and cells other than adipocytes. The authors obtained 6.9 x 10 nonruptured adipocytes from 1 g of suctioned adipose tissue; 30 percent of the original adipocytes appeared to have been ruptured. Both the XTT and glycerol-3-phosphate dehydrogenase assays provided good correlations between the number of viable adipocytes and resulting values, but only the glycerol-3-phosphate dehydrogenase assay was strictly specific for adipocytes. The ratio of adipose stromal cells to adipocytes was found to be much larger than previously described.
  
  
  CONCLUSIONS
  Single use or a combination of the viability assays used in this study can appropriately determine the number of adipocytes and other cells, although it remains difficult to assess original cells directly without tissue dissociation.
authors.affiliation: Tokyo and Yokohama, Japan From the Department of Plastic Surgery, University of Tokyo School of Medicine, and the Division of Research and Development, Biomaster, Inc.
authors.forename: Hirotaka
authors.initials: H
authors.lastname: Suga
authors.forename: Daisuke
authors.initials: D
authors.lastname: Matsumoto
authors.forename: Keita
authors.initials: K
authors.lastname: Inoue
authors.forename: Tomokuni
authors.initials: T
authors.lastname: Shigeura
authors.forename: Hitomi
authors.initials: H
authors.lastname: Eto
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Aoi
authors.forename: Harunosuke
authors.initials: H
authors.lastname: Kato
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Abe
authors.forename: Kotaro
authors.initials: K
authors.lastname: Yoshimura
mesh_terms: D017667:Adipocytes / Q000166:cytology*; D000273:Adipose Tissue / Q000166:cytology* / Q000601:surgery; D002452:Cell Count; D002470:Cell Survival; D002478:Cells, Cultured; D005260:Female; D006801:Humans; D015187:Lipectomy