Objectives: Adipose tissue is one of the many sources of regenerative cells known for many years now. Since the invention of freshly isolated SVF cells have great potential to enhance regeneration compared to lack of viability and retention of the conventional lipotransfer, SVF and SVF+Fat Grafting has become a widely used option in many branches of medicine.

Introduction: SVF cells are a complex cell fraction settled in the adipose tissue. Since its a complete fraction, there are different cells in and focused to isolate the mesenchymal cells which are adhered with protein bounds to the ECM. Due to regulatory restraints many groups have turned into create mechanical dissociation methods to isolate the mesenchymal cells. Enzyme involvement protocols are still to be considered to be the gold standard in-vitro. But so?

Materials / method: Manually harvested lipoaspirate used in this study. In First group collagenase is used with the condensated adipose tissue. Condensation was done in order to get rid of the tumescent solution and red blood cells and a small amount of triglycerids. Lipocondensation was done at 1500G for 8 mins of centrifugation.
In second group it was studied along with the use of Microlyzer blades to obtain the nanofat structure right after the same lipocondensation was performed. In the third group, SVF was obtained without the use of any enzymes and/or any incubation process.

Results: The cells were analysed using a fluoroscence microscope in order to identify nucleated cell population. The cells were then characterized according to ISCT and IFATS joint statement with cell surface markers. CD 45 negative cells were first selected, CD 34, CD 90, CD 73, CD 105 were analysed on fresh samples for characterization. Also the viability were performed with 7-AAD cell marker with high rate of viability (>85-90%) were achieved on all experimental points. While total numbers are higher with Enzymes, CD 90 and CD 73 cells were significantly better in percentage compared to enzymatic.

Conclusion: Further studies are needed to conclude the superiority of mechanical isolation of Stromal Vascular Fraction either safety or efficacy outcomes compared to the enzymatic isolation methods of Stromal Vascular Fraction.

These findings have shown that the Microlyzer device & Non-enzymatic SVF protocol offers a great yield capacity with great viability in safe conditions as the protocol does not require any enzyme which offering risk-free of enzymatic activity within safety and regulation point of view.

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