ONE S.T.E.P. TECHNIQUETM IS EFFICIENT IN HARVESTING MESENCHYMAL STEM CELLS FROM HUMAN ADIPOSE TISSUE

Background Conventional (mechanical) laser-assisted liposuction (LAL) produce an intense photothermal effect that induces adipocyte destruction. Absorption of lasers varies in tissues by their different composition (fat, water, hemoglobin), and strongly depends on different laser wavelengths along with used dosimetric parameters. The 1210-nm wavelength lasers are described to have great affinity for human adipose tissue due to tissue´s high optical absorption coefficient at 1210 nm. Safety of LAL is well documented but few studies described the effect of LAL on human mesenchymal stem cells from adipose tissue (ASCs) and much less using 1210-nm lasers. Here we compare ASCs harvested by LAL with the those harvested by One S.T.E.P. techniqueTM - Selective Tissue Engineering Photostimulation (diode laser 1210 nm), which does not cause photo-thermal lipolysis effect in contrast to conventional laser-assisted method. Methods This study was approved by the Ethic Committee of the Institution. Fat tissue was obtained from 4 patients by traditional liposuction and from 4 patients in whom One S.T.E.P. techniqueTM was used. Cells were grown in DMEM with 10% FBS and antibiotics (100 µg/mL streptomycin,100 UI/mL penicillin) at 37°C in 5% CO2 atmosphere. ASCs were characterized measuring specific antigen markers by flow cytometry. In vitro differentiation capability into osteoblasts, adipocytes, and chondrocytes were performed using defined media and evaluated by Alizarin Red, Oil red O and Alcian blue staining, respectively. Results Cells isolated from adipose tissue by both methods adhered to plastic and had a fibroblastoid morphology. Cells expressed CD29, CD90 and CD105 antigens, and lacked expression of CD14, CD34, CD45, CD80, CD117 and HLA-DR. These findings are in accordance with the definition of mesenchymal stem cells as defined by the ISCT. Moreover, cells were able to differentiate in chondrogenic, osteogenic, and adipogenic lineages under appropriated conditions. Conclusion Conventional laser lipolysis destroys both adipocytes and MSC through intense photothermal effect. The One STEP techniqueTM does not rely on lipolysis, and releases both preserved adipocytes and MSCs. In fact, here the use of One S.T.E.P techniqueTM for isolation of ASCs was shown to be an effective method to harvest ASCs cells, preserving their mesenchymal stem cell characteristics. Therefore, this method is potentially suitable for obtaining ASCs for using in regenerative medicine.