A novel method for banking dental pulp stem cells.
Dental pulp stem cells (DPSC), a cell type of mesenchymal origin showing high proliferation and plasticity, are an emerging source of adult stem cells offering interesting features in view of potential applications in regenerative medicine. These features prompted us to develop a new method to cryopreserve DPSC inside a whole tooth, thus avoiding the need to purify the cells before cryopreservation and reducing the initial costs and workload of tooth banking. In this study we cryopreserved 4 human deciduous whole teeth after digging micro-channels into the tooth with an Nd:YAG laser beam (laser piercing) to allow the cryopreservative to reach the dental pulp and preserve the cells at -80°C. Then, we isolated, expanded and characterized in vitro the stem cells after tooth thawing and mechanical fracture. In parallel, we characterized cells extracted from 2 teeth cryopreserved without laser piercing and from 4 non cryopreserved, non laser pierced, freshly fractured teeth. Our data demonstrate that DPSC isolated from laser pierced cryopreserved teeth show mesenchymal stem cells morphology, immunophenotype, viability and proliferation rate similar to those of cells isolated from fresh, non cryopreserved teeth, whereas significant loss of cell viability and proliferation rate was shown by cells isolated from teeth cryopreserved without laser piercing. These data support the use of this method for prospective whole tooth banking.
Department of Surgical Sciences, Reconstruction and Diagnostics, University of Milan, Faculty of Dentistry, IRCCS Fondazione Ca' Granda Ospedale Maggiore Policlinico, via Francesco Sforza 35, 20122 Milano, Italy.
Silvia
S
Gioventù
Gabriella
G
Andriolo
Ferruccio
F
Bonino
Stefania
S
Frasca
Lorenza
L
Lazzari
Elisa
E
Montelatici
Franco
F
Santoro
Paolo
P
Rebulla
D002454:Cell Differentiation / Q000502:physiology; D048708:Cell Growth Processes / Q000502:physiology; D015925:Cryopreservation / Q000379:methods*; D003782:Dental Pulp / Q000166:cytology*; D006801:Humans; D013234:Stem Cells / Q000166:cytology*; D014094:Tooth, Deciduous / Q000166:cytology*