Fibroblast cell proliferation assay

Fibroblast cells from a 38-year-old patient (family 600, II-18), a 40-year-old age-matched control subject (Ctrl1), and a 61-year-old male control subject (Ctrl3) at different passage numbers were used for the proliferation assay. Cells were diluted to 20,000 cells/ml with fibroblast medium and seeded into a 96-well flat bottom plate coated with gelatin and incubated at 37°C for 2 h before scanning. The plate was placed into the IncuCyte ZOOM system (Essen BioScience Inc., Ann Arbor, Michigan) and scanned under the phase 4× object with a 2-h interval. Collected data were analyzed by using the IncuCyte ZOOM’s confluence processing analysis tool (basic analysis).