Free lipid and computerized determination of adipocyte size

Abstract

The size distribution of adipocytes in a suspension, after collagenase digestion of adipose tissue, can be determined by computerized image analysis. Free lipid, forming droplets, in such suspensions implicates a bias since droplets present in the images may be identified as adipocytes. This problem is not always adjusted for and some reports state that distinguishing droplets and cells is a considerable problem. In addition, if the droplets originate mainly from rupture of large adipocytes, as often described, this will also bias size analysis. We here confirm that our ordinary manual means of distinguishing droplets and adipocytes in the images ensure correct and rapid identification before exclusion of the droplets. Further, in our suspensions, prepared with focus on gentle handling of tissue and cells, we find no association between the amount of free lipid and mean adipocyte size or proportion of large adipocytes.

Subcutaneous adipose tissue sclerostin is reduced and Wnt signaling is enhanced following 4‐weeks of sprint interval training in young men with obesity

“…One investigator analyzed three images from each visit and the mean adipocyte cell size derived from all three images was used as the average adipocyte size for each participant at each time point. The collagenase digestion method is a widely used methods to assess adipocyte cell size as it can easily differentiate lipid droplets based on their appearance due to the absence of a stained plasma membrane (Svensson et al, 2018). It has been validated against other methods and correlates well with total/regional adiposity (Laforest et al, 2017).…”

Adipose tissue and body composition in women six years after gestational diabetes: factors associated with development of type 2 diabetes

“…Adipose tissue was digested with collagenase (Type A; Roche, Mannheim, Germany); an adipocyte suspension was prepared, and adipocyte size was determined by computerized image analysis (Leica QWin V3; Leica Microsystems, Wetzlar, Germany). [36][37][38] Twelve random visual fields were photographed (DFC320, Leica Microsystems). Microspheres 98 μm in diameter (Dynal, Invitrogen, Oslo, Norway) served as reference.…”