Cardiomyocytes, incubation

Incubate the EB for 5 days in an incubator in a humidified atmosphere at 37°C and 5% CO. The cell clusters will further differentiate towards cardiomyocytes. 

The effect of the ATP-vesicles on cardiomyocyte preservation was also tested during chemical hypoxia induced by KCN. Adult Sprague-Dawley rats (200-250 g) were used and isolation of primary cardiomyocytes was performed according to well-established techniques (22-24). After baseline readings were obtained, the cells were incubated for 30 min with KCN. The group with ATP-vesicles had the lowest amount of enzymes released than that in the other groups (Fig. 4). 

Fig. 4. Cardiomyocyte enzyme release rates for CK and AST during chemical hypoxia is decreased by intracellular ATP delivery. Rat cardiomyocytes were incubated in KCN for a period of 30 min in the presence of either ATP-vesicles, lipid vesicles only, Mg-ATP only, or culture media (M199). ATP-vesicles provided significant protection from CK and AST release during chemical hypoxia. Significantly different from all groups as tested by ANOVA (p<0.05, /V=6)...

Effect of panaxadiol saponin (PDS) on phorbol ester induced change of protein kinase C activity in cardiomyocytes of rats was investigated[19]. Partially purified protein kinase C was incubated with PDS at concentration of 1 to 1000 U g/ml for ten minutes in vitro. The activity of kinase C was inhibited in a dose-dependent manner by PDS. In cardiomyocytes preincubated with PDS at concentration of250, 500,1000 P g/ml, respectively, for ten minutes, PMA-induced decrease of cytosol protein kinase C activity and increase of membrane protein kinase C activity were greatly inhibited in the same manner. PDS not only inhibited protein kinase C in vitro, but also inhibited activation of protein kinase C in cardiomyocytes. 

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