Title : Preliminary data of adipose stem cells and stem cells markers by flow cytometry and their use in regenerative medicine-plastic surgery
Abstract:
In the field of regenerative medicine, basic research and preclinical studies have been conducted to overcome clinical shortcomings with the use of mesenchymal stem cells. They are present in adult tissues, including bone marrow and adipose tissue. For many years, bone marrow-derived stem cells were the primary source of stem cells for tissue engineering applications. However, recent studies have shown that subcutaneous adipose tissue provides a clear advantage over other stem cell sources due to the ease with which adipose tissue can be accessed as well as the ease of isolating stem cells from harvested tissue. This cell population, termed adipose-derived stem cells (ADSCs), represents a promising approach to future cell-based therapies, such as tissue engineering and regeneration. Furthermore, these cells can be readily harvested in large numbers with low donor-site morbidity. It has been shown that human adult adipocytes have a roughly 10% turnover rate. One of the hallmark characteristics of human adipose-derived stem cells (hADSCs) is their ability to differentiate into cells of mesenchymal lineages. Although there is still no clear consensus on the antigen expression pattern that will clearly define hADSCs, a flow cytometry analysis protocol is presented from our lab. We also developed a novel protocol for the isolation of hASCs co-expressing at least 2 different mesenchymal adipose markers (CD271-APC ++ & CD14-FITC --). Finally the viability has been checked by Trypan Blue Microscopy and Flow Cytometry. Our preliminary result data shows us that the hADSCs antigen expression (> 97% in positive markers and < 1.5% in negative), was higher in all processed fat samples with collagenase with high viability values (> 96%), allows talking about the superiority of the collagenase processed fat on the ADSCs concentration over the pure fat and the unprocessed fat samples. More studies will allows us to present sufficient data for effective and safe lipo-injection of hADSCs in patients. We succeed to isolate hADSCs with a mean value of 8.10x10^6 with over 97% adipose antigen expression. ASCs have therapeutic potential in regenerative medicine and applications in tissue engineering. While the biologic properties of ASCs are not yet fully delineated, the cells are under clinical investigation in human trials for an array of diseases. ASCs have been widely studied for their immunomodulatory effects, antifibrotic, anti-apoptotic, and anti-oxidative capabilities in preclinical and human clinical trials. Additionally, the ASC secretome has demonstrated similar effectiveness in regenerative medicine applications.
