COMPARATIVE ASSESSMENT OF DIFFERENT APPROACHES FOR OBTAINING TERMINALLY DIFFERENTIATED MUSCLE CELLS
N.A. Smolina, A.Ya. Davidova, I.A. Schukina, A.V. Karpushev, A.B. Malashicheva, R.I. Dmitrieva, A.A. Kostareva 1
V.A. Almazov Federal Heart, Blood, and Endocrinology Centre, St. Petersburg;
1 e-mail: kostareva_aa@almazovcentre.ru
Relevant cell model is essential to study pathogenesis of muscle disorders. However, in the field of muscle research there is no ultimate cell line considered as a standard for
studying muscular and neuromuscular diseases. Standard cell line claimed to be well differentiated in muscle lineage, be morphological and physiological similar to mature muscle
cells and be easily genetically modified. Therefore, the goal of our study was to pick up available and fruitful cell model of muscle differentiation, that could be further applied for
examination of muscular disorder pathogenesis in vitro. We characterized human mesenchymal stem cells (MSC), mature murine muscle fibers and primary murine satellite
cells. It has been shown that MSC have very small capacity to myogenic differentiation; moreover, they were able to differentiate only in presence of C2C12 cells. Lentiviral
transduction exhibited rather high toxic effect on primary myofibers, and positively transduced cells were not able to response to electrical stimulation, i. e. were functionally inactive.
Satellite cells turned out to be the most fruitful cell model since they were easily transduced via lentiviruses and rapidly formed myotubes in differentiation media. Functional analysis
of obtained myotubes has confirmed their ability to react to electrical and chemical stimulations; besides, potassium and calcium channels availability has been also demonstrated
via patch-clump technique. Taken together, these results imply that satellite cells are the most promising cell line for further experiments aimed at exploring the molecular pathways
of muscle pathologies.
Key words: muscle cells, satellite cells, mesenchymal stem cells, myofibrils
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