Standardised Protocol for Establishing Primary Cell Cultures from Periodontal Fibroblasts of Healthy and Diabetic Human Subjects

Background Primary cultures of gingival and periodontal ligament fibroblasts are an important tool in research and drug development in the field of periodontologyAim To put forth a predictable protocol for harvesting primary cultures of gingival and periodontal ligament fibroblasts from healthy and diabetic subjects.Methods With prior informed consent gingival tissue and extracted teeth were collected from 20 donors following aseptic precautions. Samples were washed for two minutes with 1 povidone-iodine before transferring them to tubes containing transport medium supplemented with antibiotic-antimycotic solution. Fibroblasts were harvested from the donor tissue following the explant technique. The complete growth medium used in our study comprised of Dulbeccorsquos Modified Eagles Medium 10 foetal bovine serum and 1 antibiotic-antimycotic solution.Results The results of this study indicate a higher success rate in healthy subjects 100 compared with diabetic subjects 70 and for gingival fibroblasts 90 compared with periodontal ligament fibroblasts 80. The main reason for failure of cell culture was yeast contamination. On average gingival fibroblasts reached 70-80 confluence at 15.6 days in healthy subjects and 16 days in diabetic subjects whereas periodontal ligament fibroblasts reached confluence at 19.6 days and 21 days respectively.Conclusion The simple protocol described in this article can be used to harvest periodontal and gingival fibroblasts from healthy as well as diabetic subjects with good yield and viability.