OR26. Dedifferentiated Fat Cells versus Adipose Derived Mesenchymal Stem Cells Experimental Study about Adipose-derived Stem Cells for Cardiomyocyte Precursor using Traditional and Insert Culture

Abstract

Abstract Background and Aims The discovery of dedifferentiated fat cell (DFAT) raises the question, are AMSC and DFAT the same type of mesenchymal stem cell (MSC)? We aim to analyze the immunocytochemistry (ICC) profile of both cells and the correlation between different methods with different passage to GATA-4 and cTnT expression in vitro. Methods and Results Human subcutaneous-fat tissue was isolated and divided into, first group, using single filtration by 100 µm cell strainer and second group using double filtration with addition of 70 µm strainer after the 100 µm. Cell obtained from both groups were labeled AMSC and DFAT respectively. Prior to passaging, CD34, CD45, CD90, and CD105 were analyze by ICC. After passaging, passage 1 and 4 was exposed to Gibco PSC Cardiomyocyte Differentiation Kit for 21 days. Colonies from days 7, -14, and -21 were analyze by ICC for GATA-4 and cTnT expression. Post hoc analysis shown that there is no significance difference in MSC markers expression between groups. There were significance differences in GATA-4 expression on all observation days, which decreased significantly from the 14th to the 21st day. Statistic significancy also found on cTnT expression, with a significant increase from day 14 to day 21. The filter variable consistently has a significant effect on the quantity of GATA-4 and cTnT expressions. Conclusion The study shows that different culture methods, presumably the use of filters, will have a significant effect on the differentiation of AMSC and DFAT into cardiomyocytes.