Isolation of Human Adult Dental Pulp Stem Cells Using Enzymatic Digestion

Abstract

Objectives: The aim of the study was to culture dental pulp stem cells from non- carious, extracted human adult permanent teeth using enzymatic digestionMaterials and Methods: Human adult extracted teeth were collected from the Oral Surgery Department OPD, from the Ziauddin Dental College. Pulp was extirpated via barbed broach and isolated via enzymatic digestion using trypsin. The stem cells were cultured under conditioned media and a growth curve was plotted against the population doubling time. Cell viability was tested and finally the cells were cryopreserved for future use. Data was presented descriptively by graphical representation. Results: The stem cells exhibited spindle shaped morphology similar to mesenchymal stem cells after two weeks of isolation. The cells reached 80% confluency after a total of four weeks after which before they were processed for further assays. The growth characteristics were analyzed by calculation of population doubling time and cell viability. Conclusion: In the study successfully demonstrated the ability to isolate and culture dental pulp stem cells (DPSCs) from non-carious, extracted human adult permanent teeth using an enzymatic digestion method. The enzymatic digestion method of harvesting stem cell resulted in a stable DPSC cell growth. This method is convenient and a reliable source of stem cells for future personalized medicine research.