Harvest of superficial layers of fat with a microcannula and isolation of adipose tissue-derived stromal and vascular cells

Aesthet Surg J. 2014 May 1;34(4):601-13. doi: 10.1177/1090820X14528000. Epub 2014 Mar 31.

Abstract

Background: Adipose tissue is a source of stromal and vascular cells suitable for regenerative medical applications. Cell recovery depends on several factors, including the characteristics of the cannula used to harvest tissue.

Objectives: The authors assess whether aspiration of superficial layers of adipose tissue performed with a microcannula, rather than a standard cannula, allows for improved isolation of stromal and vascular cells, and they evaluate the angiogenic potential of the isolated cells in vitro and in vivo.

Methods: Adipose-derived stromal and stem cells (ADSC) were collected from the lipoaspirate of the abdomen and hip regions of 6 healthy female donors. For adipose tissue harvest, several options were compared: (1) a rounded-tip cannula with a length of 170 mm, a diameter of 3 mm, and a single elliptic suction port on the side near its distal end (port diameter: 3 × 9 mm) or (2) a rounded-tip infiltration cannula with a length of 170 mm, a diameter of 2 mm, and 5 round ports placed spirally along the sides of the distal cannula shaft (each port diameter: 1 mm) (Shipper Medical Technologies Corporation, Centennial, Colorado). Isolated cells were characterized for (1) expression of the endothelial specific marker CD31 by immunohistochemical and cytofluorimetric analyses and (2) tubular-like structure formation using a 3-dimensional angiogenesis assay on Matrigel. Human ADSC were transduced to express firefly luciferase as a marker suitable for bioluminescent tracking and transplantation studies into immunosuppressed mice were performed.

Results: ADSC yield was determined to be significantly higher in samples collected with the microcannula (P = .04). Moreover, isolated cells acquired typical endothelial-like morphology in vitro, formed capillary-like structures, and expressed the distinctive endothelial cell marker CD31. Cells implanted into immunosuppressed mice persisted for several weeks in areas undergoing neovascularization.

Conclusions: These results suggest that aspiration of adipose tissue with a microcannula can be a minimally invasive method to obtain clinically relevant numbers of stromal and vascular cells useful for autologous transplant procedures and for promoting tissue regeneration and neovascularization.

Keywords: adipose-derived stromal cells; autologous fat transfer; endothelial cells; lipoaspirate; liposuction; microcannula; research; stromal vascular fraction; tissue regeneration.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adipose Tissue / cytology*
  • Adipose Tissue / transplantation
  • Animals
  • Biomarkers / metabolism
  • Catheters*
  • Cell Proliferation
  • Cell Separation / methods*
  • Cell Survival
  • Cells, Cultured
  • Endothelial Cells / physiology*
  • Endothelial Cells / transplantation
  • Equipment Design
  • Female
  • Graft Survival
  • Humans
  • Lipectomy / instrumentation*
  • Mice
  • Miniaturization
  • Neovascularization, Physiologic
  • Stromal Cells / physiology*
  • Stromal Cells / transplantation
  • Time Factors
  • Tissue and Organ Harvesting / instrumentation*
  • Tissue and Organ Harvesting / methods
  • Transduction, Genetic

Substances

  • Biomarkers