Figure 2

Test of culture conditions for cardiac differentiation of hMSCs. (A) Scheme of experiment on cardiac differentiation of aspirate hMSCs in vitro. (B) PCR from human heart tissue and aspirate hMSCs used in 8 differentiation protocols (see A) demonstrating the highest efficacy of two of them that led to the increase in expression of Nkx2.5 gene along with MYH7B gene expression: 3D, DC and 2D, A/T+IMDM. (C) Clusterization of cell cultures based on the level of MYH7B, Nkx2.5, MEF2A, and MEF2D gene expression. The cell cultures tested in this experiment formed two statistically different (P = 0.036, ANOSIM) clusters at Euclidian distance of about 2.2. Note that the threedimensional cell culture in DC (3D DC) and the two-dimensional culture in IMDM pretreated with 5-azacytidine and trichostatin A (2D A/T + IMDM) formed a separate cluster. (2D - two-dimensional culture; 3D - three-dimensional culture; A/T - pretreatment with 5-azacytidin and trichostatin A; DC - cells cultured in differentiation cocktail; NCM - normal culture medium; IMDM - NCM based on Iscove's Modified Dulbecco's Medium; Differentiation cocktail - NCM based on DMEM-LG with components of differentiation cocktail).