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Simulated microgravity does not alter myogenesis gene expression in C2C12 cells
Author(s) -
Shimkus Kevin L,
Zanello Susana B,
Emami Kamal,
Wu Honglu
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.668.7
Subject(s) - myogenesis , microcarrier , c2c12 , apoptosis , microbiology and biotechnology , gene expression , myocyte , biology , cell culture , andrology , chemistry , gene , genetics , medicine
Limited work to date has been aimed at the effects of gravity (or the lack thereof) on myogenesis; the process of proliferation, differentiation, and fusion of muscle satellite cells into multinucleated myofibers. Using a bioreactor designed to mimic microgravity (0g), gene expression of key factors of myogenesis was compared between 0g‐grown cells and Earth's 1g controls. C2C12 cells were cultured on collagen microcarriers and placed in either dynamic vessels to simulate 0g or stationary vessels for the 1g control. The dynamic vessels were rotated in the horizontal plane to place the cells in freefall for 0, 1, 3, 5, or 7 days, while the control vessels remained stationary. At the appropriate times the cells were extracted, separated from the collagen beads, and processed for analysis. An aliquot of live cells was analyzed using an apoptotic assay to determine cell viability. The remaining cells were lysed for gene expression analysis via qRT‐PCR. The apoptosis assay showed no significant difference in viability between 1g and 0g for the duration of the study, and both groups showed increases in the prevalence of early stage apoptosis with time. The gene expression analysis evidenced a similar time course profile in both the static and dynamic groups. In conclusion, gravity does not appear to have a major impact on myogenesis‐relevant gene expression within the first 7 days of culture. Funded by NASA & The Huffines Institute