Cultured slow vs. fast skeletal muscle cells differ in physiology and responsiveness to stimulation

Abstract

In vitro studies have used protein markers to distinguish between myogenic cells isolated from fast and slow skeletal muscles. The protein markers provide some support for the hypothesis that satellite cells from fast and slow muscles are different, but the data are equivocal. To test this hypothesis directly, three-dimensional skeletal muscle constructs were engineered from myogenic cells isolated from fast tibialis anterior (TA) and slow soleus (SOL) muscles of rats and functionality was tested. Time to peak twitch tension (TPT) and half relaxation time (RT(1/2)) were approximately 30% slower in constructs from the SOL. The slower contraction and relaxation times for the SOL constructs resulted in left shift of the force-frequency curve compared with those from the TA. Western blot analysis showed a 60% greater quantity of fast myosin heavy chain in the TA constructs. 14 days of chronic low-frequency electrical stimulation resulted in a 15% slower TPT and a 14% slower RT(1/2), but no change in absolute force production in the TA constructs. In SOL constructs, slow electrical stimulation resulted in an 80% increase in absolute force production with no change in TPT or RT(1/2). The addition of cyclosporine A did not prevent the increase in force in SOL constructs after chronic low-frequency electrical stimulation, suggesting that calcineurin is not responsible for the increase in force. We conclude that myogenic cells associated with a slow muscle are imprinted to produce muscle that contracts and relaxes slowly and that calcineurin activity cannot explain the response to a slow pattern of electrical stimulation.

Effect of MicroRNA Modulation on Bioartificial Muscle Function

“…22,[26][27][28][29][30] The forces observed in our BAM constructs with the C2C12 cell line were on the same order of magnitude as others have reported for primary cells, 22,29,30 as there were no direct comparisons using a pure myoblast cell line. Constructs cultured with primary human myoblasts in a collagen gel had passive forces (without electrical stimulation) in the 0.5 mN range, 22 which were slightly higher than our passive forces.…”

“…Passive forces in myooids cultured with C2C12 myoblasts and 10T 1/2 fibroblasts were found to be 0.37 AE 0.05 mN, neonatal rat constructs had forces 0.31 AE 0.03 mN, and adult rat soleus myooids forces were lower at 0.12 AE 0.03 mN. 28 Engineered skeletal muscle, cultured from slow soleus rat and fast tibialis anterior rat myoblasts up to 28 days with 14 days of electrical stimulation (five electrical pulses every 4 s at 20 Hz), produced higher forces in soleus muscle (0.55 vs. 0.3 mN for no electrical stimulation), 29 which was in the same range as our results. Rat myoblasts cast into a fibrin gel produced constructs that exhibited maximum twitch and tetanic forces on the order of 0.3 and 0.8 mN, respectively.…”

Current opportunities and challenges in skeletal muscle tissue engineering

“…Therefore, frequently cell lines are used such as C2C12, which are satellite cells of C3H mice (Dennis et al, 2001;Borschel et al, 2004;Levenberg et al, 2005;Riboldi et al, 2005;Huang et al, 2006a;Boontheekul et al, 2007;Matsumoto et al, 2007). Also, satellite cells harvested from the soleus muscle of rats are used for TE of muscle tissue (Dennis and Kosnik, 2000;Beier et al, 2004;Stern-Straeter et al, 2005;Das et al, 2006;Borschel et al, 2006;Larkin et al, 2006;Bach et al, 2006;Huang et al, 2006b;Boontheekul et al, 2007). Other muscles harvested for TE research include the latissimus dorsi and rectus femoris of rats (Kamelger et al, 2004), the flexor digitorum brevis of rats (De Coppi et al, 2005), the tibialis anterior of rats (Dennis et al, 2001;Huang et al, 2005;Boontheekul et al, 2007), the extensor digitorum longus of mice (Dennis et al, 2001), the human masseter (Lewis et al, 2000;Sinanan et al, 2004;Shah et al, 2005;Stern-Straeter et al, 2008;Brady et al, 2008a) and brachioradialis (Alessandri et al, 2004) and the iliofibularis from female Xenopus laevis frogs (Jaspers et al, 2006) (Table 2).…”

“…In other words, the source of muscle fibre type predisposes satellite cells differentiation: Huang et al (2006b) showed that characteristics of muscle fibres tissue engineered from soleus muscle satellite cells are different from those obtained from tibialis anterior muscle satellite cells. For example, the contraction/relaxation time is slower in muscle constructs derived of satellite cells of soleus muscle compared to muscle constructs derived of satellite cells of tibialis anterior muscle.…”

“…It has been found that the expression of desmin, MHC and MRFs can be strongly influenced by chronic electrical stimulation, which imitates in vivo neuronal activity during myogenesis in vitro (Powell et al, 2002;Pedrotty et al, 2005). Huang et al (2006b) used this concept on muscle fibre constructs from soleus and tibialis anterior muscle satellite cells and found an increase in force production of 61-80% in soleus muscle constructs, without an increase in total MHC. They hypothesized that the increase in force production is caused by reorganization of sarcomeres within muscle fibres, promoted by chronic low-frequency electrical stimulation and/or a change in matrix produced by muscle cells, which allowed for better transmission of force to the anchor materials specifically designed to serve as artificial tendons.…”