Mechanically Isolated Stromal Vascular Fraction Provides a Valid and Useful Collagenase-Free Alternative Technique: A Comparative Study.
BACKGROUND
The use of stromal vascular fraction and adipose-derived stromal cells in tissue regeneration is now being increasingly investigated, and studies have demonstrated that adipose-derived stromal cells present differentiation and immunomodulatory capacities. The development of a rapid, inexpensive, and enzyme-free technique to isolate adipose-derived stromal cell-enriched stromal vascular fraction is a major goal for stem cell therapy. Therefore, the authors compared innovative mechanical procedures to the gold standard technique, collagenase digestion.


METHODS
Stromal vascular fraction was prepared from 21 liposuctions using either enzymatic digestion or two different mechanical methods: high vortexing/centrifugation and dissociation by intersyringe processing. The effects of tissue processing on cell count, viability, proliferation, clonogenic enrichment, and the phenotypes of the different native cell were determined. Adipose-derived stromal cell phenotypes from the different protocols, and their differentiation and immunosuppressive potential, were compared.


RESULTS
Enzymatic digestion isolated more viable cells than dissociation by intersyringe processing and vortexing/centrifugation. The expansion rate and clonogenic enrichment were higher for stromal vascular fraction isolated with collagenase. The proportion of adipose-derived stromal cells was higher in stromal vascular fraction extracted with dissociation than with enzymatic digestion and vortexing/centrifugation (p < 0.01). Interestingly, all cultured adipose-derived stromal cells displayed similar differentiation and immunosuppressive capacities.


CONCLUSIONS
Enzymatic digestion extracts more adipose-derived stromal cells, but intersyringe dissociation enables the rapid extraction of adipose-derived stromal cell-enriched stromal vascular fraction. Moreover, mechanical methods enable adipose-derived stromal cell isolation with stemness and immunosuppressive properties, similar to enzymatic digestion. Such mechanical procedures could allow easier and more rapid isolation of adipose-derived stromal cell-enriched stromal vascular fraction for practitioners.


METHODS
Therapeutic, V.
Toulouse, Rennes, and Limoges, France.|From the Department of Plastic, Reconstructive Aesthetic and Burns Surgery, University Hospital Rangueil; UMR5273 STROMALab Université Toulouse, CNRS, EFS, INSERM U1031; the Department of Plastic, Reconstructive and Aesthetic Surgery, Hospital Sud, SITI Laboratory and INSERM U917 University of Rennes 1; and the Department of Maxillofacial Surgery, University Hospital Limoges.
Benoit
B
Chaput
Toulouse, Rennes, and Limoges, France.|From the Department of Plastic, Reconstructive Aesthetic and Burns Surgery, University Hospital Rangueil; UMR5273 STROMALab Université Toulouse, CNRS, EFS, INSERM U1031; the Department of Plastic, Reconstructive and Aesthetic Surgery, Hospital Sud, SITI Laboratory and INSERM U917 University of Rennes 1; and the Department of Maxillofacial Surgery, University Hospital Limoges.
Nicolas
N
Bertheuil
Toulouse, Rennes, and Limoges, France.|From the Department of Plastic, Reconstructive Aesthetic and Burns Surgery, University Hospital Rangueil; UMR5273 STROMALab Université Toulouse, CNRS, EFS, INSERM U1031; the Department of Plastic, Reconstructive and Aesthetic Surgery, Hospital Sud, SITI Laboratory and INSERM U917 University of Rennes 1; and the Department of Maxillofacial Surgery, University Hospital Limoges.
Marina
M
Escubes
Toulouse, Rennes, and Limoges, France.|From the Department of Plastic, Reconstructive Aesthetic and Burns Surgery, University Hospital Rangueil; UMR5273 STROMALab Université Toulouse, CNRS, EFS, INSERM U1031; the Department of Plastic, Reconstructive and Aesthetic Surgery, Hospital Sud, SITI Laboratory and INSERM U917 University of Rennes 1; and the Department of Maxillofacial Surgery, University Hospital Limoges.
Jean-Louis
JL
Grolleau
Toulouse, Rennes, and Limoges, France.|From the Department of Plastic, Reconstructive Aesthetic and Burns Surgery, University Hospital Rangueil; UMR5273 STROMALab Université Toulouse, CNRS, EFS, INSERM U1031; the Department of Plastic, Reconstructive and Aesthetic Surgery, Hospital Sud, SITI Laboratory and INSERM U917 University of Rennes 1; and the Department of Maxillofacial Surgery, University Hospital Limoges.
Ignacio
I
Garrido
Toulouse, Rennes, and Limoges, France.|From the Department of Plastic, Reconstructive Aesthetic and Burns Surgery, University Hospital Rangueil; UMR5273 STROMALab Université Toulouse, CNRS, EFS, INSERM U1031; the Department of Plastic, Reconstructive and Aesthetic Surgery, Hospital Sud, SITI Laboratory and INSERM U917 University of Rennes 1; and the Department of Maxillofacial Surgery, University Hospital Limoges.
Jérôme
J
Laloze
Toulouse, Rennes, and Limoges, France.|From the Department of Plastic, Reconstructive Aesthetic and Burns Surgery, University Hospital Rangueil; UMR5273 STROMALab Université Toulouse, CNRS, EFS, INSERM U1031; the Department of Plastic, Reconstructive and Aesthetic Surgery, Hospital Sud, SITI Laboratory and INSERM U917 University of Rennes 1; and the Department of Maxillofacial Surgery, University Hospital Limoges.
Nicolas
N
Espagnolle
Toulouse, Rennes, and Limoges, France.|From the Department of Plastic, Reconstructive Aesthetic and Burns Surgery, University Hospital Rangueil; UMR5273 STROMALab Université Toulouse, CNRS, EFS, INSERM U1031; the Department of Plastic, Reconstructive and Aesthetic Surgery, Hospital Sud, SITI Laboratory and INSERM U917 University of Rennes 1; and the Department of Maxillofacial Surgery, University Hospital Limoges.
Louis
L
Casteilla
Toulouse, Rennes, and Limoges, France.|From the Department of Plastic, Reconstructive Aesthetic and Burns Surgery, University Hospital Rangueil; UMR5273 STROMALab Université Toulouse, CNRS, EFS, INSERM U1031; the Department of Plastic, Reconstructive and Aesthetic Surgery, Hospital Sud, SITI Laboratory and INSERM U917 University of Rennes 1; and the Department of Maxillofacial Surgery, University Hospital Limoges.
Luc
L
Sensebé
Toulouse, Rennes, and Limoges, France.|From the Department of Plastic, Reconstructive Aesthetic and Burns Surgery, University Hospital Rangueil; UMR5273 STROMALab Université Toulouse, CNRS, EFS, INSERM U1031; the Department of Plastic, Reconstructive and Aesthetic Surgery, Hospital Sud, SITI Laboratory and INSERM U917 University of Rennes 1; and the Department of Maxillofacial Surgery, University Hospital Limoges.
Audrey
A
Varin
D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D002478:Cells, Cultured; D002498:Centrifugation; D017364:Collagenases; D006801:Humans; D015187:Lipectomy; D059630:Mesenchymal Stem Cells / Q000502:physiology; D010641:Phenotype; D050507:Subcutaneous Fat, Abdominal / Q000166:cytology*
D017364:Collagenases