CHARACTERISATION OF AN IN VITRO AGED CELL CULTURE MODEL
MARKO BUNC 1, SARA M HANNING 2, JOSE M PRIETO 3, JULIJANA KRISTL 1, MINE ORLU 3*
1University of Ljubljana, Faculty of Pharmacy, Aškerčeva cesta 7, 1000 Ljubljana, Slovenia
2University of Auckland, School of Pharmacy, Private Bag 92019, Auckland 1142, New Zealand
3University College London, UCL School of Pharmacy, 29-39 Brunswick Square, London WC1N 1AX, United Kingdom
*corresponding author: m.orlu@ucl.ac.uk
Download Full Article

Abstract:
The present study aims to provide a detailed protocol to obtain an aged cell culture model, which is conveniently
characterised in terms of morphology, metabolism, and key biochemical markers of senescence. Commercial Human Dermal
Fibroblasts (HDF) (67-year-old female Asian donor) were cultured over a span of 35 passages to determine at what passages
they express the phenotypes found in vivo in the geriatric population. The growth rate and density (counting, Trypan Blue) at
confluence declined 4- and 3-fold, respectively. During 15 passages in vitro ageing the fibroblast metabolic capacity (XTT)
was heavily reduced (by 75%), while conversely the content of cellular proteins (SRB) slightly increased (2 fold). Senescent
cells (SA-β-gal positive) were continuously emerging from 0 to 20%. Freezing/thawing at this stage restored metabolic
performance to original values which declined at a similar rate as before. Passages 5 - 10 of our selected aged HDFs were
found to mimic in vivo tissue of older adults to the greatest extent.

Top