The aim of this study is to check hypothesis that increase of fluorescence at 460 nm emission upon myoblast differentiation display alteration of NADH quantity that change NAD+/NADH ratio and could be a myoblast marker of entering into differentiation process.

To evaluate NAD(P)H influence on total cell fluorescence emission intensity at 460 nm wavelenght were used metabolism effectors: carbonyl cyanide 3-chlorophenylhydrazone (CCCP) to obtain minimal NADH fluorescence; and KCN to obtain maximal NADH fluorescence. Assuming, that CCCP caused maximal oxidation and that KCN caused maximum reduction, estimated that NAD+/NADH ratio decreased upon myoblast differentiation. CCCP effect showed that about 95 % of myoblast fluorescence at 460 nm emission is dependent on NAD(P)H. The fluorescence intensity at 460 nm was increasing with myoblast differentiation. Using fluorescence measuring estimated average NADH concentration significantly increased on 10 day of differentiation compared with control cells.

Changes of fluorescence intensity at 535 nm reveal accumulation of FAD upon myoblast differentiation. Therefore, the best indicator of myoblast differentiation stage is ratio of fluorescence emission at 535 and 460 nm.

Our results indicate that quick, quantitative and real-time fluorescence method may be useful for non-invasive detection and evaluation of myoblast differentiation stage.

Raktažodžiai: satellite cell, NADH fluorescence, myoblast differentiation