期刊
IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL
卷 49, 期 2, 页码 114-121出版社
SPRINGER
DOI: 10.1007/s11626-012-9576-y
关键词
Mouse dental papilla cell; Reversible immortalization; Tamoxifen; CRE; Odontoblast
资金
- National Natural Science Foundation of China [30872880, 81070797]
- National 973 project of China [2010CB534915]
- Wuhan University [20103040101000162]
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.
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