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S143 Premature Ageing And Skeletal Muscle Dysfunction In Copd Patients: Development Of A Cell Culture Model
  1. R Lahkdar1,
  2. G Choudhury1,
  3. J McLeish1,
  4. EM Drost1,
  5. L McGlynn2,
  6. PG Shiels2,
  7. W MacNee1,
  8. RA Rabinovich1
  1. 1Edinburgh Lung and the Environment Group Initiative (ELEGI), Centre for Inflammation and Research, Queens Medical Research Institute, Edinburgh, Edinburgh, UK
  2. 2University of Glasgow, College of Medical, Veterinary and Life Sciences Institute of Cancer Sciences, Glasgow, UK

Abstract

Introduction COPD is a disease of accelerated ageing, as increased cellular-ageing (senescence) occurs in the lungs of these patients. We aim at developing a primary skeletal muscle cell culture (Human skeletal muscle satellite cells [HSKMC]) model of muscle ageing to study the cascade of events that occur in muscle senescence in vitro and to explore the effect of inflammation (TNF alpha) and oxidative stress (H2O2), two of the putative mechanisms related to muscle dysfunction and/wasting, on muscle differentiation and on protein loss in differentiated cells.

Methods and results HSKMC were cultured to senescence when the cells stopped replicating. DNA was isolated from cells in serial passages of culture. Telomere length, a marker of biological ageing, was measured by qPCR and expressed as the ratio of telomere repeat copy number to single gene copy number in the experimental sample relative to a control sample (relative T/S ratio) (n = 3). Preliminary results show a progressive shortening of telomere length with cellular ageing when comparing early (passage 2, 1.433 ± 0.05 relative T/S ratio) with a later passage (passage 15, 0.340 ± 0.2 relative T/S ratio) (Fig 1).

Conclusion We have developed a novel in vitro model of ageing skeletal muscle cells, which will help us to assess the role of accelerated ageing in muscle dysfunction and/wasting in COPD patients.

Dr Lakhdar was funded by an LTERS fellowship grant.

Abstract S143 Figure 1

Telomere Length analysis in HSKMC

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