Metabolic Remodeling During H9c2 Myoblast Differentiation: Relevance for In Vitro Toxicity Studies
H9c2 cells, derived from the ventricular part of an E13 BDIX rat heart, possess a proliferative and relatively undifferentiated phenotype but can be readily directed to differentiate under reduced serum conditions originating cells presenting muscle features. Skeletal or cardiac phenotypes can be originated depending on whether or not serum reduction is accompanied by a daily treatment with all-trans-retinoic acid. In the present study, we aimed to characterize and compare the metabolic profile of H9c2 cells at various differentiation states, correlating the differences between different populations with muscle-specific development. We determined that H9c2 myoblasts remodel their metabolism upon differentiation, with undifferentiated cells more reliant on glycolysis, as demonstrated by higher lactate production rates. Differentiated cells adopted a more oxidative metabolism with better coupling between the glycolytic and oxidative pathways, which is indicative of a metabolic evolvement toward a higher energetic efficiency state. Our findings emphasize the metabolic differences between differentiated and undifferentiated H9c2 cells and raise caution on how to adequately select the H9c2 differentiation state that will act as the better model for the design of experimental studies.
KeywordsH9c2 myoblasts Cell differentiation Metabolism NMR isotopomer analysis Cardiotoxicology
The authors thank Gonçalo Pereira, Dr. Mario Grãos and Dr. Anabela Rolo for their assistance in the different phases of the work. Financial Support: SLP is the recipient of a PhD fellowship (SFRH/BD/37933/2007). The present work was funded by research grants POCI/SAU-OBS/55802/2004, PTDC/QUI/64358/2006, and PTDC/EBB-EBI/101114/2008 from the Portuguese Foundation for Science and Technology. We acknowledge the National NMR Network for access to the facilities. The Varian VNMRS 600 MHz spectrometer is part of the National NMR Network and was purchased in the framework of the National Program for Scientific Re-equipment, contract REDE/1517/RMN/2005, with funds from POCI 2010 (FEDER) and Fundação para a Ciência e a Tecnologia (FCT).
Conflict of interest
- 14.Saeedi, R., Saran, V. V., Wu, S. S., Kume, E. S., Paulson, K., Chan, A. P., et al. (2009). AMP-activated protein kinase influences metabolic remodeling in H9c2 cells hypertrophied by arginine vasopressin. American Journal of Physiology Heart Circulatory Physiology, 296, H1822–H1832.CrossRefGoogle Scholar
- 19.Mukhopadhyay, P., Rajesh, M., Batkai, S., Kashiwaya, Y., Hasko, G., Liaudet, L., et al. (2009). Role of superoxide, nitric oxide, and peroxynitrite in doxorubicin-induced cell death in vivo and in vitro. American Journal of Physiology Heart Circulatory Physiology, 296, H1466–H1483.CrossRefGoogle Scholar
- 21.Liu, Z., Song, X. D., Xin, Y., Wang, X. J., Yu, H., Bai, Y. Y., et al. (2009). Protective effect of chrysoeriol against doxorubicin-induced cardiotoxicity in vitro. Chinese Medicine Journal, 122, 2652–2656. (Engl).Google Scholar