IADR Abstract Archives
Characterization of Dental-tissue Derived Mesenchymal Stromal Cells in Animal Component-free Culture System
Objectives: The use of animal components such as fetal bovine serum (FBS) during the isolation and culture of human dental mesenchymal stromal cells (DSCs) is one of the major obstacles that hinders clinical translation for stem cell-based therapies. Here, we aim to isolate and culture the DSCs from extracted tooth (dental pulp and periodontal ligament) under complete animal component-free (ACF) culture system and investigate its impact on proliferation ability, surface marker expression and tri-lineage differentiation capacity.
Methods: The dental pulp and periodontal ligament were isolated from healthy donors aged 15 to 30 years. DSCs were isolated through explant culture in DMEM medium +10% FBS, and ACF medium. For culture in ACF system, the DSCs were transferred to culture plates coated with MesencultTM-ACF attachment substrate, cultured in MesencultTM-ACF Plus medium and subcultured using ACF dissociation reagents. The proliferation potential, cryopreservation, surface marker expression (classic MSC markers: CD73, CD90, CD105; MHC antigens: Class I,II; neural-lineage markers: CD56, CD271), tri-lineage differentiation potential of the cells under ACF conditions were investigated.
Results: The DSCs cultured in ACF media showed high proliferative ability compared to FBS-containing medium. More than 95% cells showed a positive expression of classic MSC surface markers. A positive expression of MHC Class I and a negative HLA Class II expression was observed. The DSCs also expressed the neural-lineage marker CD56. Maintenance of proliferation ability, surface marker expression and tri-lineage differentiation after cryopreservation was noted in ACF system. The DSCs exhibited a higher tri-lineage differentiation ability in ACF system compared to those cultured in FBS-containing medium.
Conclusions: The DSCs cultured in ACF conditions demonstrated a higher proliferative ability, positive stem cell surface marker expression and higher tri-lineage differentiation potential. Hence, such comprehensive ACF culture approach will aid towards culture of clinical-grade DSCs for future stem cell-based therapies.
Methods: The dental pulp and periodontal ligament were isolated from healthy donors aged 15 to 30 years. DSCs were isolated through explant culture in DMEM medium +10% FBS, and ACF medium. For culture in ACF system, the DSCs were transferred to culture plates coated with MesencultTM-ACF attachment substrate, cultured in MesencultTM-ACF Plus medium and subcultured using ACF dissociation reagents. The proliferation potential, cryopreservation, surface marker expression (classic MSC markers: CD73, CD90, CD105; MHC antigens: Class I,II; neural-lineage markers: CD56, CD271), tri-lineage differentiation potential of the cells under ACF conditions were investigated.
Results: The DSCs cultured in ACF media showed high proliferative ability compared to FBS-containing medium. More than 95% cells showed a positive expression of classic MSC surface markers. A positive expression of MHC Class I and a negative HLA Class II expression was observed. The DSCs also expressed the neural-lineage marker CD56. Maintenance of proliferation ability, surface marker expression and tri-lineage differentiation after cryopreservation was noted in ACF system. The DSCs exhibited a higher tri-lineage differentiation ability in ACF system compared to those cultured in FBS-containing medium.
Conclusions: The DSCs cultured in ACF conditions demonstrated a higher proliferative ability, positive stem cell surface marker expression and higher tri-lineage differentiation potential. Hence, such comprehensive ACF culture approach will aid towards culture of clinical-grade DSCs for future stem cell-based therapies.