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Controlled Heat Stress Promotes Myofibrillogenesis During Myogenesis
Samuel Boas BS, Anil Chaturvedi MS, Corinne Wee MD, Anand Kumar MD
University Hospitals Cleveland Medical Center
2019-02-09
Presenter: Samuel Boas
Affidavit:
I certify that the majority of the work is the project of the medical student.
Director Name: Anand Kumar MD
Author Category: Medical Student
Presentation Category: Basic Science Research
Abstract Category: General Reconstruction
Significance: Hyperthermic treatment of cellular tissues is an emerging modality for biomedical applications. Specific effects of the timing and temperature of heat stress on muscle regeneration after injury remains understudied. This study aimed to characterize high myogenic capacity Muscle Derived Stem Cells (MDSCs) response to hyperthermic therapy.
Methods: MDSCs from murine hind limb tissue were isolated using a standardized isolation protocol using Type 1 collagen for cell sorting. MDSCs (1x 106/well) from pre plate 3/4 were cultured and expanded to confluence at different temperatures intervals (37, 39, 41°C). Myotube differentiation was quantified at specific times (1,3,5 days) using immunofluorescent cell staining. Statistical analysis was preformed using SPSS.
Results: MDSCs demonstrated significant changes in morphology based on temperature and temporal changes. Early transient moderate and severe hypothermia promoted myotuble growth. MDSCs cultured at 39°C and 41°C grew significantly longer at 48 hours (515, 478 mm) compared with control myotubules cultured at 37°C (339 mm)(p<.001, p<.001). Prolonged severe hypothermia was deleterious to muscle growth and expansion. At 72 hours, MDSCs cultured at 41°C had a significantly lower nuclei density (1454 nuclei/mm2) than MDSCs cultured at 37°C (1890 nuclei/mm2)(p=.008), and after five days, MDSCs cultured at 41°C (803 nuclei/mm2) had significantly lower nuclei density than MDSCs cultured at 37°C (1800 nuclei/mm2) and 39°C (1624 nuclei/mm2)(p<.001, p<.001).
Conclusion: MDSCs treated with early/transient (<72 hours) moderate hyperthermia (39°C) demonstrated significant improvement in myotube growth. Persistent and severe hyperthermia significantly decreased muscle growth. Future studies quantifying cellular processes through pathway-focused high-throughput gene expression profiling are warranted.
