Differentiation of human adipose-derived stem cells towards cardiomyocytes is facilitated by laminin

Adipose-derived stein cells (ASCs) are promising candidates for therapy in myocardial infarction (MI). However. the frequency of human ASCs that differentiate towards cardiomyocytes is low. We hypothesized that adherence to extracellular matrix molecules that are upregulated after MI might increase human stem cell differentiation towards cardiomyocytes. We analysed putative ASC differentiation oil fibronectin-coated, laminin-coated and uncoated Culture plates. Expression of cardiac marker,, in cells was analysed 1, 3 and 5 weeks after stimulation with 5-aza-2-deoxycytidine. After I week, mRNA expression of myosin light chain-2 alpha (MLC-2 alpha), all early marker in cardiomyocyte development, was increased significantly in treated cells, independent of coating. At 5 weeks, however, mRNA expression of the late cardiomyocyte development marker SERCA2 alpha was only significantly increased in 5-aza-2-deoxycytidine-treated cells Cultured oil laminin. Significantly higher numbers of cells were immunopositive for MLC-2 alpha in cultures of treated cells grown oil laminin-coated wells, when compared with cultures of treated cells grown Oil uncoated wells, both at I week and at 5 weeks. Furthermore, after 3 weeks, significantly more alpha-actinin- and desmin-positive cells were detected after treatment with 5-aza-2-deoxycytidine, but only ill uncoated wells. After 5 weeks, however, the number of desmin-positive cells was only significantly increased after treatment of cells with 5-aza-2-deoxycytidine and culture oil laminin (61% positive cells). Thus, we have found that a high percentage of human ASCs call be differentiated towards cardiomyocytes; this effect call be improved by laminin, especially during late differentiation.