Establishment and Characterization of a Tamoxifen-mediated Reversible Immortalized Mouse Dental Papilla Cell Line

Overview

Journal In Vitro Cell Dev Biol Anim

Publisher Springer

Specialties Biology
Cell Biology

Date 2013 Jan 10

PMID 23299318

Citations 12

Authors

Heng Lin

Huan Liu

Qin Sun

Guohua Yuan

Lu Zhang

Zhi Chen

Affiliations

Soon will be listed here.

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.

Citing Articles

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