Figure 1
Paracrine effects of MSCs on Dox-induced oxidative stress and apoptosis of neonatal rat cardiomyocytes (NRCMs) in-vitro.
a) 1 × 104 iPSC-MSCs or BM-MSCs were seeded on 0.3 μm pore size transwell inserts and 1 × 104 NRCMs were seeded on the bottom of a 96-well culture plate with or without presence of 1 μM Dox. 24 hours later, the transwell inserts were removed and MTT assay was used to measure NRCMs cell viability (i). The percentage of cell viability of each group was calculated from triple experiments (ii) (*p < 0.01 vs. control group; #p < 0.01 vs. Dox group; †p < 0.01 vs. BM-MSCs-CdM group). b,c) NRCMs were treated with or without MSCs-CdM (BM-MSCs-CdM or iPSC-MSCs-CdM) under challenge of 1 μM Dox for 24 hours. In living cell culture conditions, H2DCFDA (b-i) and TUNEL (c-i) staining were used to measure ROS generation and cell apoptosis respectively. Quantitative measurement of ROS (b-ii) and apoptotic rate (c-ii) was counted from five viewing fields in each group and triple experiments were performed (*p < 0.01 vs. control group; #p < 0.01 vs. Dox group; †p < 0.01 vs. BM-MSCs-CdM group). The concentration of MDA (b-iii) among the different groups was also measured (*p < 0.01 vs. control group; #p < 0.01 vs. Dox group; †p < 0.01 vs. BM-MSCs-CdM group). d) The effect of iPSC-MSCs-CdM showed a dose-dependent manner in attenuation of ROS generation and apoptosis induced by Dox. The concentration of MDA (d-i) and apoptosis (d-ii) among the different groups were also measured among the different groups was measured (*p < 0.01 vs. control group; # p < 0.01 vs. Dox group; †p < 0.01 vs. iPSC-MSCs-CdM-10 μl group; $p < 0.01 vs. iPSC-MSCs-CdM-20 μl group).
