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Characterization of primary human skeletal muscle cells from multiple commercial sources.

Owens J, Moreira K, Bain G - In Vitro Cell. Dev. Biol. Anim. (2013)

Bottom Line: Primary human skeletal muscle cells have recently become available from a number of commercial vendors.However, only limited characterization of these cells has been reported to date.Finally, the myotubes were efficiently infected with recombinant adenovirus, providing a tool for genetic modification.

View Article: PubMed Central - PubMed

Affiliation: Tissue Repair Research Unit, Pfizer, 200 Cambridge Park Drive, Cambridge, MA, 02140, USA, Jane.Owens@Pfizer.com.

ABSTRACT
There is a significant unmet need for safe, anabolic muscle therapies to treat diseases and conditions associated with severe muscle weakness and frailty. The identification of such therapies requires appropriate cell-based screening assays to select compounds for further development using animal models. Primary human skeletal muscle cells have recently become available from a number of commercial vendors. Such cells may be valuable for studying the mechanisms that direct muscle differentiation, and for identifying and characterizing novel therapeutic approaches for the treatment of age- and injury-induced muscle disorders. However, only limited characterization of these cells has been reported to date. Therefore, we have examined four primary human muscle cell preparations from three different vendors for their capacity to differentiate into multinucleated myotubes. Two of the preparations demonstrated robust myotube formation and expressed characteristic markers of muscle differentiation. Furthermore, these myotubes could be induced to undergo morphological atrophy- and hypertrophy-like responses, and atrophy could be blocked with an inhibitor of myostatin signaling, a pathway that is known to negatively regulate muscle mass. Finally, the myotubes were efficiently infected with recombinant adenovirus, providing a tool for genetic modification. Taken together, our results indicate that primary human muscle cells can be a useful system for studying muscle differentiation, and may also provide tools for studying new therapeutic molecules for the treatment of muscle disease.

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Effect of TWEAK on HSMM and SkMDC. Representative images of MYH2/Hoechst-labeled myotubes from cultures differentiated for 4 d in differentiation medium with TWEAK (1 μg/ml). (Magnification ×10; scale bar = 100 microns).
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Fig3: Effect of TWEAK on HSMM and SkMDC. Representative images of MYH2/Hoechst-labeled myotubes from cultures differentiated for 4 d in differentiation medium with TWEAK (1 μg/ml). (Magnification ×10; scale bar = 100 microns).

Mentions: In addition to Mstn and Dex, the cytokine TNF-related weak inducer of apoptosis (TWEAK) has been reported to be a potent inducer of muscle atrophy (Dogra et al. 2007a) and blocks the differentiation of C2C12 myoblasts into myotubes (Dogra et al. 2006, 2007b). In our studies, treatment of HSMM and SkMDC myotubes with TWEAK had no observable effects (data not shown). However, when added to the culture at the start of myotube differentiation, the differentiation of SkMDC was completely blocked, while it had no effect on HSMM differentiation (Fig. 3). Expression of the TWEAK receptor, FN14, has been demonstrated on human muscle satellite cells (Girgenrath et al. 2006), but it is possible that FN14 is not expressed by HSMM which would explain the absence of an effect of TWEAK in these cultures. We examined FN14 expression in both cell populations by real-time RT-PCR and found that it was expressed in both myoblasts and myotubes and showed no changes during differentiation (data not shown).Figure 3.


Characterization of primary human skeletal muscle cells from multiple commercial sources.

Owens J, Moreira K, Bain G - In Vitro Cell. Dev. Biol. Anim. (2013)

Effect of TWEAK on HSMM and SkMDC. Representative images of MYH2/Hoechst-labeled myotubes from cultures differentiated for 4 d in differentiation medium with TWEAK (1 μg/ml). (Magnification ×10; scale bar = 100 microns).
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3824271&req=5

Fig3: Effect of TWEAK on HSMM and SkMDC. Representative images of MYH2/Hoechst-labeled myotubes from cultures differentiated for 4 d in differentiation medium with TWEAK (1 μg/ml). (Magnification ×10; scale bar = 100 microns).
Mentions: In addition to Mstn and Dex, the cytokine TNF-related weak inducer of apoptosis (TWEAK) has been reported to be a potent inducer of muscle atrophy (Dogra et al. 2007a) and blocks the differentiation of C2C12 myoblasts into myotubes (Dogra et al. 2006, 2007b). In our studies, treatment of HSMM and SkMDC myotubes with TWEAK had no observable effects (data not shown). However, when added to the culture at the start of myotube differentiation, the differentiation of SkMDC was completely blocked, while it had no effect on HSMM differentiation (Fig. 3). Expression of the TWEAK receptor, FN14, has been demonstrated on human muscle satellite cells (Girgenrath et al. 2006), but it is possible that FN14 is not expressed by HSMM which would explain the absence of an effect of TWEAK in these cultures. We examined FN14 expression in both cell populations by real-time RT-PCR and found that it was expressed in both myoblasts and myotubes and showed no changes during differentiation (data not shown).Figure 3.

Bottom Line: Primary human skeletal muscle cells have recently become available from a number of commercial vendors.However, only limited characterization of these cells has been reported to date.Finally, the myotubes were efficiently infected with recombinant adenovirus, providing a tool for genetic modification.

View Article: PubMed Central - PubMed

Affiliation: Tissue Repair Research Unit, Pfizer, 200 Cambridge Park Drive, Cambridge, MA, 02140, USA, Jane.Owens@Pfizer.com.

ABSTRACT
There is a significant unmet need for safe, anabolic muscle therapies to treat diseases and conditions associated with severe muscle weakness and frailty. The identification of such therapies requires appropriate cell-based screening assays to select compounds for further development using animal models. Primary human skeletal muscle cells have recently become available from a number of commercial vendors. Such cells may be valuable for studying the mechanisms that direct muscle differentiation, and for identifying and characterizing novel therapeutic approaches for the treatment of age- and injury-induced muscle disorders. However, only limited characterization of these cells has been reported to date. Therefore, we have examined four primary human muscle cell preparations from three different vendors for their capacity to differentiate into multinucleated myotubes. Two of the preparations demonstrated robust myotube formation and expressed characteristic markers of muscle differentiation. Furthermore, these myotubes could be induced to undergo morphological atrophy- and hypertrophy-like responses, and atrophy could be blocked with an inhibitor of myostatin signaling, a pathway that is known to negatively regulate muscle mass. Finally, the myotubes were efficiently infected with recombinant adenovirus, providing a tool for genetic modification. Taken together, our results indicate that primary human muscle cells can be a useful system for studying muscle differentiation, and may also provide tools for studying new therapeutic molecules for the treatment of muscle disease.

Show MeSH
Related in: MedlinePlus