Abstract

Objectives: The aim of the project consisted of the control of viability and phenotype of adipose derived cells using a guided procedure with the two types of cannulas having 0.8 mm and 1 mm side port holes and processed following SEFFI (Superficial Enhanced Fluid Fat Injection) technique; the results were compared with tissue harvested with a standard liposuction technique and processed with a standard procedure consisting of enzymatic digestion (collagenase). . Introduction: Injection of autologous adipose-derived stem cells (ADSCs) and a stromal vascular fraction (VSF) into dermal and subdermal layers promises regenerative advantages by improving skin volume and rejuvenation. It's extremely important to set a Good Protocol (GP) to harvest and manipulate the adipose tissue in order to achieve the higher cell viability and the highest amount of SVF cells and ADSCs. Materials / method: This study was performed on adipose tissues harvested from 7 patients (6 females and 1 male) with an average age of 48.5 years with 3 different techniques. Adipose tissue using a 0.8- and 1-mm cannula were monitored for their cell viability using absorbance spectroscopy ; a.t. harvested using Coleman was processed by enzymatic treatment as quality control of tissue. For phenotype analysis Cells were incubated with antibodies anti human against CD90-FITC, CD73-PeCy7, CD44-PE, CD31-PECy5, CD235a- PECy7, CD34-FITC, CD45-FITC and CD146-FITC and read using the flow cytometer . Results: Lipoaspirate tissue derived by 0.8- and 1 mm cannula from all samples proved to be vital and possess viable cells.Average of the absorbance was similar immediately after plating (t0) and 72 hours after (t72) for the two cannulas, 0.8- and 1 mm cannula. The two systems proved to equally harvest vital tissue. Morphologically, cells harvested by different cannula or isolated by enzymatic digestion showed differences in their morphology. Conclusion: The results proved the vitality of adipose tissue harvested using the different cannulas.Tissue was vital in both cannulas and the values were comparable to the isolation of stromal vascular fraction (SVF) by enzymatic digestion, the gold standard procedure for adipose tissue derived cells. Cells to be defined mesenchymal stem cells, need to adhere to plastic surface and express specific surface antigens, especially CD44, CD90 and CD73.Cells derived from each individual were able to attach and proliferate for at least three passages in culture and show the typical fibroblastic morphology.