In vitro exercise model using contractile human and mouse hybrid myotubes

Weijian Chen,1 Mazvita R. Nyasha,1 Masashi Koide,2 Masahiro Tsuchiya,3 Naoki Suzuki,4 Yoshihiro Hagiwara,2 Masashi Aoki,4 and Makoto Kanzaki
Source: Scientific Reports
Publication Date: (2019)
Issue: 9: 11914
Research Area:
Basic Research
Cells used in publication:
Skeletal Muscle Myoblast, (HSMM) human
Species: human
Tissue Origin: skeletal muscle

Researchers used Electro=pulse stimulation to study the repsonse of human and mouse hybrid myotubes In vitro. Previous studies using only human cells to measure contractility and other produce dmixed results. The hybrid model proved to be much more effective.


Contraction of cultured myotubes with application of electric pulse stimulation (EPS) has been utilized for investigating cellular responses associated with actual contractile activity. However, cultured myotubes derived from human subjects often exhibit relatively poor EPS-evoked contractile activity, resulting in minimal contraction-inducible responses (i.e. myokine secretion). We herein describe an "in vitro exercise model", using hybrid myotubes comprised of human myoblasts and murine C2C12 myoblasts, exhibiting vigorous contractile activity in response to EPS. Species-specific analyses including RT-PCR and the BioPlex assay allowed us to separately evaluate contraction-inducible gene expressions and myokine secretions from human and mouse constituents of hybrid myotubes. The hybrid myotubes, half of which had arisen from primary human satellite cells obtained from biopsy samples, exhibited remarkable increases in the secretions of human cytokines (myokines) including interleukins (IL-6, IL-8, IL-10, and IL16), CXC chemokines (CXCL1, CXCL2, CXCL5, CXCL6, CXCL10), CC chemokines (CCL1, CCL2, CCL7, CCL8, CCL11, CCL13, CCL16, CCL17, CCL19, CCL20, CCL21, CCL22, CCL25, CCL27), and IFN-? in response to EPS-evoked contractile activity. Together, these results indicate that inadequacies arising from human muscle cells are effectively overcome by fusing them with murine C2C12 cells, thereby supporting the development of contractility and the resulting cellular responses of human-origin muscle cells. Our approach, using hybrid myotubes, further expands the usefulness of the "in vitro exercise model".