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Phenotypic Analysis of Stromal Vascular Fraction after Mechanical Shear Reveals Stress-Induced Progenitor Populations

Chaput, Benoit M.D.; Laloze, Jerome M.D.; Grolleau, Jean-Louis M.D.; Espagnolle, Nicolas Ph.D.; Bertheuil, Nicolas M.D.; Varin, Audrey Ph.D.

Plastic & Reconstructive Surgery: April 2017 - Volume 139 - Issue 4 - p 1024e–1025e
doi: 10.1097/PRS.0000000000003202
Letters

Department of Plastic, Reconstructive, and Aesthetic Surgery, and, STROMAlab, UMR5273 CNRS/UPS/EFS-INSERM U1031, Rangueil Hospital, Toulouse, France

Department of Plastic, Reconstructive, and Aesthetic Surgery, Rangueil Hospital, Toulouse, France

STROMAlab, UMR5273 CNRS/UPS/EFS-INSERM U1031, Rangueil Hospital, Toulouse, France

STROMAlab, UMR5273 CNRS/UPS/EFS-INSERM U1031, Rangueil Hospital, Toulouse, France, Department of Plastic, Reconstructive, and Aesthetic Surgery, Hospital Sud, University of Rennes 1, Rennes, France, SITI Laboratory, Etablissement Français du Sang Bretagne, Rennes University Hospital, Rennes, France

Department of Plastic, Reconstructive, and Aesthetic Surgery, and, STROMAlab, UMR5273 CNRS/UPS/EFS-INSERM U1031, Rangueil Hospital, Toulouse, France

Correspondence to Dr. Chaput, Department of Plastic and Reconstructive Surgery, Rangueil University Hospital, Avenue du Professeur Jean Poulhes, 31 000 Toulouse, France, benoitchaput31@gmail.com

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Sir:

We read with great interest the article entitled “Phenotypic Analysis of Stromal Vascular Fraction after Mechanical Shear Reveals Stress-Induced Progenitor Populations” by Banyard et al.1 This is a new article focusing on the stromal vascular fraction obtained after intersyringe processing developed by Tonnard et al.2 Development of a new and efficient technique of stromal vascular fraction and fat preparation is currently a hot topic in both fat grafting and regenerative medicine.

The intersyringe processing allows preparation of two distinct products. The first is the unfiltered emulsified fat, which contains mainly dead adipocytes and extracellular matrix. This whitish product, which has a paste aspect, can be used for volume defects but requires a minimum of 25-gauge needles for injection because of the extracellular matrix that can block finer needles. Banyard et al. analyzed this product, which is different from nanofat graft. Indeed, the nanofat grafting product is filtered before use to remove the remaining connective tissue. Here, the authors have filtered after enzymatic digestion, which does not reflect the nano fat grafting. The use of collagenase before filtration will therefore yield a completely different cell population. In their Discussion, the authors are aware of it but, clinically, this cannot be the same use, as several times we experienced that this product does not pass through needles smaller than 24- to 25-gauge.

The second product obtained after intersyringe processing and filtration is a mechanically isolated stromal vascular fraction, which differs from the one obtained after collagenase digestion. Indeed, we recently analyzed this second product obtained after the Tonnard procedure without collagenase digestion, and we obtained different results compared with Banyard et al.3,4 Indeed, even when we used different phenotypic markers, we demonstrated that stromal vascular fraction obtained after intersyringe processing contains significantly fewer hematopoietic, endothelial, and red blood cells (CD235a+ cells) than stromal vascular fraction obtained after enzymatic digestion, and we conclude that this technique produces an adipose-derived stem cell–enriched stromal vascular fraction (579,195 adipose-derived stem cells/g versus 47,859 adipose-derived stem cells/g; p < 0.001). The reduction in cell viability is a problem we also found, and the work of Osinga et al. was somewhat surprising.5 Thus, we noted twofold less viable stromal vascular fraction after mechanical digestion compared with enzymatic stromal vascular fraction digestion.

Finally, the authors concluded that intersyringe processing increases the number of CD105+/SSEA3+ muse cells in the stromal vascular fraction. However, muse cells exhibit characteristics of both mesenchymal and pluripotent stem cells. They are double-positive for CD105 and stage-specific embryonic antigen-3, but they also express pluripotency markers, including Oct3/4, Nanog, and Sox2, and are capable of differentiating into cells of ectodermal, endodermal, and mesodermal lineages.6 It would be necessary to test the differentiation capacities and the expression of pluripotency markers of these cells before concluding that intersyringe processing induces multilineage differentiating stress-enduring cells. Then, the authors conclude that there is a significant up-regulation of the mesenchymal stem cell population in stress-induced stromal vascular fraction compared with control stromal vascular fraction. However, the CD34+/CD45 cell population consists of not just mesenchymal stem cells but also contains endothelial cells.7 Moreover, it would be interesting to determine the progenitor capacities (colony-forming units) of these enduring stress cells compared with conventional adipose-derived stem cells obtained after enzymatic digestion.

We congratulate the authors for this study that opens new perspectives regarding the regenerative potential of mechanical adipose tissue processing. It seems necessary to investigate the actual regenerative potential of these stress-induced progenitor obtained after intersyringe processing.

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DISCLOSURE

The authors have no financial interest to declare in relation to the content of this communication.

Benoit Chaput, M.D.

Jerome Laloze, M.D.

Department of Plastic, Reconstructive, and

Aesthetic Surgery, and

STROMAlab, UMR5273 CNRS/UPS/EFS-INSERM U1031

Rangueil Hospital

Toulouse, France

Jean-Louis Grolleau, M.D.

Department of Plastic, Reconstructive, and

Aesthetic Surgery

Rangueil Hospital

Toulouse, France

Nicolas Espagnolle, Ph.D.

STROMAlab, UMR5273 CNRS/UPS/EFS-INSERM U1031

Rangueil Hospital

Toulouse, France

Nicolas Bertheuil, M.D.

STROMAlab, UMR5273 CNRS/UPS/EFS-INSERM U1031

Rangueil Hospital

Toulouse, France

Department of Plastic, Reconstructive, and

Aesthetic Surgery

Hospital Sud

University of Rennes 1

Rennes, France

SITI Laboratory

Etablissement Français du Sang Bretagne

Rennes University Hospital

Rennes, France

Audrey Varin, Ph.D.

Department of Plastic, Reconstructive, and

Aesthetic Surgery, and

STROMAlab, UMR5273 CNRS/UPS/EFS-INSERM U1031

Rangueil Hospital

Toulouse, France

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REFERENCES

1. Banyard DA, Sarantopoulos CN, Borovikova AA, et alPhenotypic analysis of stromal vascular fraction after mechanical shear reveals stress-induced progenitor populations. Plast Reconstr Surg. 2016;138:237e–247e.
2. Tonnard P, Verpaele A, Peeters G, Hamdi M, Cornelissen M, Declercq HNanofat grafting: Basic research and clinical applications. Plast Reconstr Surg. 2013;132:1017–1026.
3. Chaput B, Bertheuil N, Escubes M, et alMechanically isolated stromal vascular fraction provides a valid and useful collagenase-free alternative technique: A comparative study. Plast Reconstr Surg. 2016;138:807–819.
4. Bertheuil N, Chaput BA novel and effective strategy for the isolation of adipose-derived stem cells: Minimally manipulated adipose-derived stem cells for more rapid and safe stem cell therapy. Plast Reconstr Surg. 2015;135:454e–455e.
5. Osinga R, Menzi NR, Tchang LA, et alEffects of intersyringe processing on adipose tissue and its cellular components: Implications in autologous fat grafting. Plast Reconstr Surg. 2015;135:1618–1628.
6. Kuroda Y, Kitada M, Wakao S, et alUnique multipotent cells in adult human mesenchymal cell populations. Proc Natl Acad Sci USA 2010;107:8639–8643.
7. Bertheuil N, Chaput B, Ménard C, et alAdipose-derived stromal cells: History, isolation, immunomodulatory properties and clinical perspectives (in French). Ann Chir Plast Esthet. 2015;60:94–102.
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