Rebekah Yu, Shyam Kolvekar, Daryll Baker, George Hamilton, David Abraham, Janice Tsui
Division of Surgery and Interventional Science, University College London, London, United Kingdom.
OBJECTIVES: Patients with critical limb ischemia (CLI) have attributable myopathy that impairs their functional ability as well as dampening the positive effect of successful revascularization outcomes. Adult skeletal muscle retains the potential to repair following different injury patterns. We aim to investigate the potential differences in proliferation and migration capability of human myoblasts with CLI and asymptomatic control patients.
METHODS: Gastrocnemius muscle biopsies were obtained from patients with critical limb ischemia and control samples were obtained from non-ischemic patients. Human myoblasts were isolated and cultured from each sample and stained with the myoblast marker desmin. Confirmed myoblast cultures (<5% alternative cell type contamination) were then used to investigate the proliferative capability (MTT assay) as well as migratory potential (scratch-wound assay) under both ischemic and normoxic conditions of each group.
RESULTS: Myoblasts isolated from CLI patients demonstrated greater proliferative ability in comparison to control samples. However, control samples revealed greater capacity to heal scratch wounds made in a monolayer of myoblasts in both normoxia and hypoxic conditions.
CONCLUSIONS: Isolation of human myoblasts, especially from disease groups, provides a useful platform to perform in-vitro analysis to better understand and characterise pathology. CLI myoblasts exhibited enhanced proliferative but diminished migratory potential in comparison to control myoblasts.
AUTHOR DISCLOSURES: D. Abraham: Nothing to disclose; D. Baker: Nothing to disclose; G. Hamilton: Nothing to disclose; S. Kolvekar: Nothing to disclose; J. Tsui: Nothing to disclose; R. Yu: Nothing to disclose.
Posted April 2013