Reply: Mechanical Isolation of Stromal Vascular Fraction

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We read with interest the comment made by Cedric Ménard about our article entitled “Mechanically Isolated Stromal Vascular Fraction Provides a Valid and Useful Collagenase-Free Alternative Technique: A Comparative Study.”1 Macrophages are certainly the key elements of initiation and control of the inflammatory response,2 and interactions between mesenchymal stem cells and innate immune cells play an essential role in the balance between proinflammatory and antiinflammatory responses that maintain tissue integrity. Moreover, mesenchymal stem cells can modulate adaptive immune cells such as T lymphocytes through modulation of the polarization and the function of macrophages. Indeed, mesenchymal stem cells induce antiinflammatory macrophages that promote immunosuppression through T regulatory lymphocyte activation.3 Therefore, as indicated by Cedric Ménard, it would be interesting to compare the immunomodulatory properties toward macrophages of adipose-derived stem cells extracted with the enzymatic method to the one isolated with the two different mechanical techniques. However, like Kokai and Rubin,4 we think that the key question for the clinical use of these cell preparation is to determine the effect of isolation processing on their native immunosuppressive capacities. Characterization of immunosuppressive properties of freshly isolated stromal vascular fraction or adipose-derived stem cells would bring crucial information for the field of stromal cell therapy that can modify drastically the condition of their clinical use.
Ménard highlighted that stromal vascular fraction isolated with dissociation or with vortex/centrifugation techniques contains a lower amount of adipose-derived stem cells compared with enzymatic digestion and that it could be an issue for their use on cell therapy. A recent study showed that cultured adipose-derived stem cells, compared with freshly isolated stromal vascular fraction, are increasingly used in clinical study,5 and our results demonstrated that cultivated adipose-derived stem cells obtained after the intersyringe processes presented similar proliferation and immunosuppressive capacities compared to cultivated adipose-derived stem cells isolated with enzymatic digestion. Therefore, the dissociation procedure could be considered as a fast and reliable alternative method to obtain enough cultivated adipose-derived stem cells to use for laboratory experiments and for stem cell therapy. Moreover, the dissociation by intersyringe processing allows a rapid extraction of adipose-derived stem cell–enriched stromal vascular fraction (close to 40 percent of adipose-derived stem cells) containing a small proportion of contaminating cells. Therefore, the improvement of cell viability and adipose-derived stem cell yield could allow the development of an innovative stromal vascular fraction and adipose-derived stem cell isolation process that could be an efficient alternative to enzymatic digestion and that can meet the strict criteria within the regulatory framework for use in regenerative medicine. We are pleased that our article raises the interest of the scientific and medical communities and that our work can contribute to increased knowledge in the field of stromal cell therapy.
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