PDL fibroblasts were isolated from a 25-year-old healthy woman whose premolar tooth was extracted as part of orthodontic treatment. The tooth was extracted by a dental surgeon and stored in modified Eagle's medium (MEM), supplemented with 5% fetal bovine serum (FBS) (Gibco, Grand Island, NY, USA) and antibiotics, including penicillin and streptomycin (Sigma Chemical, St. Louis, Mo, USA).
Periodontal tissue was scraped off the root using a sterile scalpel and rinsed with phosphate-buffered saline (PBS) twice. The scraped tissue was digested in a mixture of 0.05% trypsin and 0.02% EDTA in Hank’s balanced salt solution with decreased calcium and magnesium salts. Fibroblasts were cultured in sterile cell culture flasks (SPL Life, Science, Gyeonggi-do, South Korea), including Dulbecco’s modified Eagle’s medium, 10% FBS, 100 IU/mL penicillin, 100 ug/mL streptomycin, and 2 mM/mL glutamine in an incubator containing 5% CO2 and 95% air at 37°C. The culture medium was refreshed every 2-3 days during the cell culture process, and the cells were passaged after one week. After the 4th passage, the cells had a spindle shape and comprised a homogenous cell population.12 (link) The cells were maintained in the culture medium under the mentioned conditions.
Periodontal tissue was scraped off the root using a sterile scalpel and rinsed with phosphate-buffered saline (PBS) twice. The scraped tissue was digested in a mixture of 0.05% trypsin and 0.02% EDTA in Hank’s balanced salt solution with decreased calcium and magnesium salts. Fibroblasts were cultured in sterile cell culture flasks (SPL Life, Science, Gyeonggi-do, South Korea), including Dulbecco’s modified Eagle’s medium, 10% FBS, 100 IU/mL penicillin, 100 ug/mL streptomycin, and 2 mM/mL glutamine in an incubator containing 5% CO2 and 95% air at 37°C. The culture medium was refreshed every 2-3 days during the cell culture process, and the cells were passaged after one week. After the 4th passage, the cells had a spindle shape and comprised a homogenous cell population.12 (link) The cells were maintained in the culture medium under the mentioned conditions.