title: Cryopreservation of periodontal ligament cells with magnetic field for tooth banking.
abstract: The purpose of this study was to establish a long-term tooth cryopreservation method that can be used for tooth autotransplantation. Human periodontal ligament (PDL) cells were frozen in 10% dimethyl sulfoxide (Me(2)SO) using a programmed freezer with a magnetic field. Cells were cryopreserved for 7 days at -150 degrees C. Immediately after thawing, the number of surviving cells was counted and the cells were cultured; cultured cells were examined after 48 h. Results indicated that a 0.01 mT of a magnetic field, a 15-min hold-time, and a plunging temperature of -30 degrees C led to the greatest survival rate of PDL cells. Based on these findings, whole teeth were cryopreserved under the same conditions for 1 year. The organ culture revealed that the PDL cells of cryopreserved tooth with a magnetic field could proliferate as much as a fresh tooth, although the cells did not appear in the cryopreserved tooth without a magnetic field. Histological examination and the transmission electron microscopic image of cryopreserved tooth with a magnetic field did not show any destruction of cryopreserved cells. In contrast, severe cell damage was seen in cells frozen without a magnetic field. These results indicated that a magnetic field programmed freezer is available for tooth cryopreservation.
authors.affiliation: Department of Orthodontics and Craniofacial Developmental Biology, Hiroshima University Graduate School of Biomedical Science, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, Japan. mkaku@hiroshima-u.ac.jp
authors.forename: M
authors.initials: M
authors.lastname: Kaku
authors.forename: H
authors.initials: H
authors.lastname: Kamada
authors.forename: T
authors.initials: T
authors.lastname: Kawata
authors.forename: H
authors.initials: H
authors.lastname: Koseki
authors.forename: S
authors.initials: S
authors.lastname: Abedini
authors.forename: S
authors.initials: S
authors.lastname: Kojima
authors.forename: M
authors.initials: M
authors.lastname: Motokawa
authors.forename: T
authors.initials: T
authors.lastname: Fujita
authors.forename: J
authors.initials: J
authors.lastname: Ohtani
authors.forename: N
authors.initials: N
authors.lastname: Tsuka
authors.forename: Y
authors.initials: Y
authors.lastname: Matsuda
authors.forename: H
authors.initials: H
authors.lastname: Sunagawa
authors.forename: R A M
authors.initials: RA
authors.lastname: Hernandes
authors.forename: N
authors.initials: N
authors.lastname: Ohwada
authors.forename: K
authors.initials: K
authors.lastname: Tanne
mesh_terms: D002470:Cell Survival / Q000187:drug effects / Q000528:radiation effects; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004574:Electromagnetic Fields; D006801:Humans; D008280:Magnetics; D046529:Microscopy, Electron, Transmission; D009924:Organ Culture Techniques; D010513:Periodontal Ligament / Q000166:cytology* / Q000187:drug effects / Q000528:radiation effects; D014015:Tissue Banks
chemical_list: D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide