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Establishment of Immortalized Mouse Bmp2 Knock‐Out Dental Papilla Mesenchymal Cells Necessary for Study of Odontoblastic Differentiation and Odontogenesis
Author(s) -
Wu Lian,
Wang Feng,
Donly Kevin J.,
Wan Chunyan,
Luo Daoshu,
Harris Stephen E.,
MacDougall Mary,
Chen Shuo
Publication year - 2015
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.25061
Subject(s) - dental papilla , odontoblast , dentinogenesis , mesenchymal stem cell , bone morphogenetic protein 2 , microbiology and biotechnology , extracellular matrix , cellular differentiation , dmp1 , biology , dentin , pathology , genetics , gene , in vitro , medicine , viral matrix protein
Bmp2 is essential for dentin formation. Bmp2 cKO mice exhibited similar phenotype to dentinogenesis imperfecta, showing dental pulp exposure, hypomineralized dentin, and delayed odontoblast differentiation. As it is relatively difficult to obtain lot of primary Bmp2 cKO dental papilla mesenchymal cells and to maintain a long‐term culture of these primary cells, availability of immortalized deleted Bmp2 dental papilla mesenchymal cells is critical for studying the underlying mechanism of Bmp2 signal in odontogenesis. In this study, our goal was to generate an immortalized deleted Bmp2 dental papilla mesenchymal (iBmp2 ko/ko dp) cell line by introducing Cre recombinase and green fluorescent protein (GFP) into the immortalized mouse floxed Bmp2 dental papilla mesenchymal (iBmp2 fx/fx dp) cells. iBmp2 ko/ko dp cells were confirmed by GFP and PCR. The deleted Bmp2 cells exhibited slow cell proliferation rate and cell growth was arrested in G2 phase. Expression of tooth‐related marker genes and cell differentiation were decreased in the deleted cells. Importantly, extracellular matrix remodeling was impaired in the iBmp2 ko/ko dp cells as reflected by the decreased Mmp‐9 expression. In addition, with exogenous Bmp2 induction, these cell differentiation and mineralization were rescued as well as extracellular matrix remodeling was enhanced. Therefore, we for the first time described establishment of iBmp ko/ko cells that are useful for study of mechanisms in regulating dental papilla mesenchymal cell lineages. J. Cell. Physiol. 9999: 2588–2595, 2015. © 2015 Wiley Periodicals, Inc.