Analysis for apoptosis and necrosis on adipocytes, stromal vascular fraction, and adipose-derived stem cells in human lipoaspirates after liposuction

Plast Reconstr Surg. 2013 Jan;131(1):77e-85e. doi: 10.1097/PRS.0b013e3182729ff7.

Abstract

Background: Adipose-derived stem cells have become the most studied adult stem cells. The authors examined the apoptosis and necrosis rates for adipocyte, stromal vascular fraction, and adipose-derived stem cells in fresh human lipoaspirates.

Methods: Human lipoaspirate (n = 8) was harvested using a standard liposuction technique. Stromal vascular fraction cells were separated from adipocytes and cultured to obtain purified adipose-derived stem cells. A panel of stem cell markers was used to identify the surface phenotypes of cultured adipose-derived stem cells. Three distinct stem cell subpopulations (CD90/CD45, CD105/CD45, and CD34/CD31) were selected from the stromal vascular fraction. Apoptosis and necrosis were determined by annexin V/propidium iodide assay and analyzed by flow cytometry.

Results: The cultured adipose-derived stem cells demonstrated long-term proliferation and differentiation evidenced by cell doubling time and positive staining with oil red O and alkaline phosphatase. Isolated from lipoaspirates, adipocytes exhibited 19.7 ± 3.7 percent apoptosis and 1.1 ± 0.3 percent necrosis; stromal vascular fraction cells revealed 22.0 ± 6.3 percent of apoptosis and 11.2 ± 1.9 percent of necrosis; stromal vascular fraction cells had a higher rate of necrosis than adipocytes (p < 0.05). Among the stromal vascular fraction cells, 51.1 ± 3.7 percent expressed CD90/CD45, 7.5 ± 1.0 percent expressed CD105/CD45, and 26.4 ± 3.8 percent expressed CD34/CD31. CD34/CD31 adipose-derived stem cells had lower rates of apoptosis and necrosis compared with CD105/CD45 adipose-derived stem cells (p < 0.05).

Conclusions: Adipose-derived stem cells had a higher rate of apoptosis and necrosis than adipocytes. However, the extent of apoptosis and necrosis was significantly different among adipose-derived stem cell subpopulations.

MeSH terms

  • Adipocytes, White / cytology
  • Adipocytes, White / pathology*
  • Adult
  • Adult Stem Cells / cytology
  • Adult Stem Cells / metabolism
  • Adult Stem Cells / pathology*
  • Antigens, CD / metabolism
  • Apoptosis*
  • Biomarkers / metabolism
  • Cell Differentiation
  • Cells, Cultured
  • Centrifugation
  • Female
  • Flow Cytometry
  • Hematopoietic Stem Cells / metabolism
  • Humans
  • Lipectomy*
  • Male
  • Mesenchymal Stem Cells / metabolism
  • Middle Aged
  • Necrosis
  • Osteoblasts / cytology
  • Phenotype
  • Subcutaneous Fat / pathology*
  • Subcutaneous Fat / surgery

Substances

  • Antigens, CD
  • Biomarkers