title: Establishment and characterization of a tamoxifen-mediated reversible immortalized mouse dental papilla cell line.
abstract: Odontoblasts are a type of non-proliferating and terminally differentiated cells that play an important role in the pulpo-dentinal complex. Mouse dental papilla cells (mDPCs), which can differentiate into odontoblast-like cells in vitro, have a limited life span. We combined the traditional strategy of "Cre/LoxP-based reversible immortalization" with a tamoxifen-regulated Cre recombination system to generate a tamoxifen-mediated reversibly immortalized mouse dental papilla cell line, mDPCET. mDPCs were sequentially transduced with a floxed SV40 T antigen-TK (SV40Tag-TK) and an (ERT2)Cre(ERT2)-expressing plasmid. Clonal-isolated SV40Tag- and Cre-positive cells showed modified growth characteristics and a significantly extended life span. When mDPCET cells were treated with 4-hydroxytamoxifen, (ERT2)Cre(ERT2) translocated from the cytoplasm to the nucleus and caused the excision of SV40Tag-TK, which led to the reversion of mDPCETs. After the immortalization was reversed, the cells underwent replicative senescence and transitioned into a more differentiated state. Tamoxifen-mediated reversible immortalization, therefore, allows for the expansion of primary mDPCs, leads to the production of odontoblast-like cells that retain most odontoblast-specific properties, and can represent a safe and ready-to-use method due to its simple manipulation.
authors.affiliation: State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST), School and Hospital of Stomatology, Wuhan University, Wuhan, China.
authors.forename: Heng
authors.initials: H
authors.lastname: Lin
authors.forename: Huan
authors.initials: H
authors.lastname: Liu
authors.forename: Qin
authors.initials: Q
authors.lastname: Sun
authors.forename: Guohua
authors.initials: G
authors.lastname: Yuan
authors.forename: Lu
authors.initials: L
authors.lastname: Zhang
authors.forename: Zhi
authors.initials: Z
authors.lastname: Chen
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D003771:Dental Papilla / Q000166:cytology* / Q000187:drug effects; D005822:Genetic Vectors; D051379:Mice; D009804:Odontoblasts / Q000166:cytology / Q000187:drug effects; D061251:Primary Cell Culture; D013629:Tamoxifen / Q000494:pharmacology*
chemical_list: D013629:Tamoxifen