A protocol for isolation and culture of mesenchymal stem cells from human gingival tissue

Human gingiva-derived mesenchymal stem cells (GMSCs) have been considered to be a better source of MSCs for cell therapy in some immunological diseases. We describe a protocol for isolation and culture of mesenchymal stem cells (MSCs) from human gingival tissue in detail, which provides a methodology to help clinical researches and clinical trial. GMSCs are generally isolated from a remnant or discarded tissue following a routine dental procedure, then cultured in complete culture medium at 37°C in a humidified tissue culture incubator with 5% CO₂ and 95% O₂. Non-adherent cells are removed after 48~72 h and the fresh medium is replaced. When primary cultures become 80%~90% confluent, the plastic-adherent cells are treated with 0.25% trypsin-EDTA and subcultured. A purified population of GMSCs can be obtained 2-3 weeks after the initiation of culture.