Objectives/Hypothesis: Normal human vocal fold fibroblast (hVFF) primary cell lines are unavailable commercially and are very difficult to acquire, subsequently little is known about their characteristics. The purpose of this study was to compare the morphological and proliferation characteristics and gene expression of hVFFs from different aged donors.
Study Design: In vitro.
Methods: We developed three normal hVFF primary cell lines from donors aged 21 (21T), 59 (59T) and 79 (79T) years. We characterized their morphological features, proliferative abilities, telomere lengths, and their functional gene expression by quantitative real-time PCR.
Results: The 21T line maintained a typical spindle shape until passage 14 whereas 59T and 79T changed morphology to wider, shorter cells at passage 7. Proliferation rates were constant for the 21T through passage 14; 59T's proliferative half-life was passage 9, whereas 79T maintained lower proliferation rates from passage 4. Gene expression levels for fibronectin, collagen I, collagen VI, procollagen I and elastin demonstrated similar patterns for all lines, however, relative amounts decreased with the age of donor. Telomere lengths did not show differences related with donor age.
Conclusions: hVFF primary cultures have limited proliferative capacity. The morphology, proliferation, differentiation, and gene expression levels of VFF can be affected by age, but senescence patterns were similar across the ages.