IADR Abstract Archives
Isolation of Mesenchymal Stem Cells From Adult and Deciduous Teeth
Objectives: The aim of this research is to extract different types of mesenchymal stem cells from teeth. Types of stem cells isolated from the tooth include Stem cells from the apical papilla (SCAP), Desiduous pulp stem cells (SHED), Dental pulp stem cells (DPSC), Periodontal ligament stem cells (PDL), Bone marrow-derived mesenchymal stem cells (BMSC), oral mesenchymal stem cell and gingiva mesenchymal stem cells (GMSC).
Methods: The mesenchymal stem cells are isolated form third molars, which are extracted in routine procedure, from patients between the age of 18-30 years old. First tissue that is isolated from the tooth is SCAP. It is isolated when tooth is not fully developed. The apical papilla that is separated from the apex with a scalpel. The DPSC are isolated from the pulp tissue. With dental bud the crown is separated from the root. Further the pulp tissue is gently separated from the crown and root for DPSC. When the root is separated from the crown and the pulp is removed, the tissue on the surface of the root with the whole root is seeded for the isolation of the periodontal ligament (PDL). SHED is isolated from deciduous teeth. BMSC type of cells are gathered with a routine extraction. GMSC were isolated from the gingiva.
The tissue is digested in a solution of 3mg/mL collagenase Type I and 4mg/ml dispase dissolved in DMEM for 1 hour at CO2 incubator. The suspensions of cells are cultured into 25 cm2 plastic flasks with DMEM supplemented with 20% fetal bovine serum (FBS, Gibco BRL), 100U/mL antibiotic/antimicotic (ThermoScinetific), for primary culture and then incubated at 37°C .and 5% CO2. When cells reached 80–90 % confluence, they were trypsinized (0.05 % trypsin-EDTA, Gibco) (5 min, 37 °C).
Results: We successfully isolated the tissue, incubated and cultivated the cells. After that we checked with specific stem cells markers.
Conclusions: With these experiments we have proven that there is a possibility to extract multipotent mesenchymal stem cell types from adult teeth. If successfully differentiated, they can be used to develop new techniques for stem cell therapy and drug development.
Methods: The mesenchymal stem cells are isolated form third molars, which are extracted in routine procedure, from patients between the age of 18-30 years old. First tissue that is isolated from the tooth is SCAP. It is isolated when tooth is not fully developed. The apical papilla that is separated from the apex with a scalpel. The DPSC are isolated from the pulp tissue. With dental bud the crown is separated from the root. Further the pulp tissue is gently separated from the crown and root for DPSC. When the root is separated from the crown and the pulp is removed, the tissue on the surface of the root with the whole root is seeded for the isolation of the periodontal ligament (PDL). SHED is isolated from deciduous teeth. BMSC type of cells are gathered with a routine extraction. GMSC were isolated from the gingiva.
The tissue is digested in a solution of 3mg/mL collagenase Type I and 4mg/ml dispase dissolved in DMEM for 1 hour at CO2 incubator. The suspensions of cells are cultured into 25 cm2 plastic flasks with DMEM supplemented with 20% fetal bovine serum (FBS, Gibco BRL), 100U/mL antibiotic/antimicotic (ThermoScinetific), for primary culture and then incubated at 37°C .and 5% CO2. When cells reached 80–90 % confluence, they were trypsinized (0.05 % trypsin-EDTA, Gibco) (5 min, 37 °C).
Results: We successfully isolated the tissue, incubated and cultivated the cells. After that we checked with specific stem cells markers.
Conclusions: With these experiments we have proven that there is a possibility to extract multipotent mesenchymal stem cell types from adult teeth. If successfully differentiated, they can be used to develop new techniques for stem cell therapy and drug development.