Human Gingival Fibroblast Isolation and Culture

Human gingival fibroblasts (HGF) were purchased from ATCC (HGF-1) and also isolated from gingival biopsies as previously described^{18 (link)–20 (link)}. All donors signed a written informed consent to participate. This study was approved by the Institutional Review Board at the University of the Pacific (protocol #17–66) and was conducted in accordance with the Helsinki Declaration of 1975, as revised in 2000. Briefly, gingival biopsies from clinically non-inflamed tissue were retrieved from the gingival margin. Primary cells were obtained by explants cultivated in Dulbecco’s Modified Eagle’s Medium^# supplemented with 10% fetal bovine serum^# and antibiotics (100 UI/mL penicillin/streptomycin)^#. HGF were maintained at 37°C in a humidified 5% CO₂ incubator and were used for experiments between the 4^th and 8^th passages.