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Cell Culture of a Stromal-Vascular Fraction and Its Image Analysis

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A stromal vascular fraction of human adipose tissue was cultivated in T25 flask up to six passages, using DMEM/Ham's F12(1:1) media containing 10% FBS, Penicillin and streptomycin at 37°C under humidified atmosphere of 95% air : 5% CO2. The SV fraction is comprised of mixture of cells, such as mature adipocytes, preadipocytes, fibroblasts, vascular smooth muscle cells and endothelial cells1, 2). In order to trace the variation of cell mixture, image analysis was applied to analyze the mixed cell population. After 7 to 10 days of cultivation, cells were fixed with 70% ethanol and stained by coomassie brilliant blue. Images were harvested from an inverted microscope with digital camera. A cell image analysis software, CellProfiler(www.cellprofiler.org), was adopted for extracting data including cell number, area, perimeter, eccentricity, solidity, extent, euler number and form factor. Parameters such as cell perimeter,
eccentricity and extent were found to be critical determinants for discrimination of the mixtures of cells in the SV-fraction.

저자정보

  • Won-Mo Kang Kangwon National University, Dept. of Bioengineering and Technology
  • Eun-Jong Kim Kangwon National University, Dept. of Bioengineering and Technology
  • Dae-cheol Jung Kangwon National University, Dept. of Bioengineering and Technology
  • Won Hur Kangwon National University, Dept. of Bioengineering and Technology

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