Immortalized Mouse Dental Papilla Mesenchymal Cells Preserve Odontoblastic Phenotype and Respond to Bone Morphogenetic Protein 2

Overview

Journal In Vitro Cell Dev Biol Anim

Publisher Springer

Specialties Biology
Cell Biology

Date 2013 Jul 2

PMID 23813243

Citations 10

Authors

Feng Wang

Li-An Wu

Wentong Li

Yuan Yang

Feng Guo

Qingping Gao

Hui-Hsiu Chuang

Lisa Shoff

Wei Wang

Shuo Chen

Affiliations

Soon will be listed here.

Abstract

Odontogenesis is the result of the reciprocal interactions between epithelial-mesenchymal cells leading to terminally differentiated odontoblasts. This process from dental papilla mesenchymal cells to odontoblasts is regulated by a complex signaling pathway. When isolated from the developing tooth germs, odontoblasts quickly lose their potential to maintain the odontoblast-specific phenotype. Therefore, generation of an odontoblast-like cell line would be a good surrogate model for studying the dental mesenchymal cell differentiation into odontoblasts and the molecular events of dentin formation. In this study, immortalized dental papilla mesenchymal cell lines were generated from the first mouse mandibular molars at postnatal day 3 using pSV40. These transformed cells were characterized by RT-PCR, immunohistochemistry, Western blot, and analyzed for alkaline phosphatase activity and mineralization nodule formation. One of these immortalized cell lines, iMDP-3, displayed a high proliferation rate, but retained the genotypic and phenotypic characteristics similar to primary cells as determined by expression of tooth-specific markers and demonstrated the ability to differentiate and form mineralized nodules. Furthermore, iMDP-3 cells had high transfection efficiency as well as were inducible and responded to BMP2 stimulation. We conclude that the establishment of the stable murine dental papilla mesenchymal cell line might be used for studying the mechanisms of dental cell differentiation and dentin formation.

Citing Articles

On-demand chlorine dioxide solution enhances odontoblast differentiation through desulfation of cell surface heparan sulfate proteoglycan and subsequent activation of canonical Wnt signaling.

Inubushi T, Priyanka N, Watanabe M, Takahashi Y, Kusano S, Kurosaka H Front Cell Dev Biol. 2023; 11:1271455.

PMID: 37954207 PMC: 10637356. DOI: 10.3389/fcell.2023.1271455.

Immortalized cell lines derived from dental/odontogenic tissue.

Zeng Y, Liu L, Huang D, Song D Cell Tissue Res. 2023; 393(1):1-15.

PMID: 37039940 DOI: 10.1007/s00441-023-03767-5.

Mesenchymal stem cells: A comprehensive methods for odontoblastic induction.

Koh B, Sulaiman N, Ismadi S, Ramli R, Yunus S, Idrus R Biol Proced Online. 2021; 23(1):18.

PMID: 34521356 PMC: 8442352. DOI: 10.1186/s12575-021-00155-7.

Changes of mitochondrial respiratory function during odontogenic differentiation of rat dental papilla cells.

Zhang F, Jiang L, He Y, Fan W, Guan X, Deng Q J Mol Histol. 2017; 49(1):51-61.

PMID: 29189956 DOI: 10.1007/s10735-017-9746-z.

Klf5 Mediates Odontoblastic Differentiation through Regulating Dentin-Specific Extracellular Matrix Gene Expression during Mouse Tooth Development.

Chen Z, Zhang Q, Wang H, Li W, Wang F, Wan C Sci Rep. 2017; 7:46746.

PMID: 28440310 PMC: 5404268. DOI: 10.1038/srep46746.

References

Lin H, Liu H, Sun Q, Yuan G, Zhang L, Chen Z . Establishment and characterization of a tamoxifen-mediated reversible immortalized mouse dental papilla cell line. In Vitro Cell Dev Biol Anim. 2013; 49(2):114-21. DOI: 10.1007/s11626-012-9576-y. View

Thesleff I, Nieminen P . Tooth morphogenesis and cell differentiation. Curr Opin Cell Biol. 1996; 8(6):844-50. DOI: 10.1016/s0955-0674(96)80086-x. View

George A, Sabsay B, Simonian P, Veis A . Characterization of a novel dentin matrix acidic phosphoprotein. Implications for induction of biomineralization. J Biol Chem. 1993; 268(17):12624-30. View

McKnight D, Hart P, Hart T, Hartsfield J, Wilson A, Wright J . A comprehensive analysis of normal variation and disease-causing mutations in the human DSPP gene. Hum Mutat. 2008; 29(12):1392-404. PMC: 5534847. DOI: 10.1002/humu.20783. View

Matsubara T, Kida K, Yamaguchi A, Hata K, Ichida F, Meguro H . BMP2 regulates Osterix through Msx2 and Runx2 during osteoblast differentiation. J Biol Chem. 2008; 283(43):29119-25. PMC: 2662012. DOI: 10.1074/jbc.M801774200. View

Feng J, Yang G, Yuan G, Gluhak-Heinrich J, Yang W, Wang L . Abnormalities in the enamel in bmp2-deficient mice. Cells Tissues Organs. 2011; 194(2-4):216-21. PMC: 3178081. DOI: 10.1159/000324644. View

Wu L, Feng J, Wang L, Mu Y, Baker A, Donly K . Immortalized mouse floxed Bmp2 dental papilla mesenchymal cell lines preserve odontoblastic phenotype and respond to BMP2. J Cell Physiol. 2010; 225(1):132-9. PMC: 2980836. DOI: 10.1002/jcp.22204. View

Galler K, Schweikl H, Thonemann B, DSouza R, Schmalz G . Human pulp-derived cells immortalized with Simian Virus 40 T-antigen. Eur J Oral Sci. 2006; 114(2):138-46. DOI: 10.1111/j.1600-0722.2006.00327.x. View

Park G, Morasso M . Bone morphogenetic protein-2 (BMP-2) transactivates Dlx3 through Smad1 and Smad4: alternative mode for Dlx3 induction in mouse keratinocytes. Nucleic Acids Res. 2002; 30(2):515-22. PMC: 99823. DOI: 10.1093/nar/30.2.515. View

10.

Dong J, Amor D, Aldred M, Gu T, Escamilla M, Macdougall M . DLX3 mutation associated with autosomal dominant amelogenesis imperfecta with taurodontism. Am J Med Genet A. 2005; 133A(2):138-41. DOI: 10.1002/ajmg.a.30521. View

11.

Linde A, Goldberg M . Dentinogenesis. Crit Rev Oral Biol Med. 1993; 4(5):679-728. DOI: 10.1177/10454411930040050301. View

12.

Ye L, Macdougall M, Zhang S, Xie Y, Zhang J, Li Z . Deletion of dentin matrix protein-1 leads to a partial failure of maturation of predentin into dentin, hypomineralization, and expanded cavities of pulp and root canal during postnatal tooth development. J Biol Chem. 2004; 279(18):19141-8. DOI: 10.1074/jbc.M400490200. View

13.

Ulsamer A, Ortuno M, Ruiz S, Susperregui A, Osses N, Rosa J . BMP-2 induces Osterix expression through up-regulation of Dlx5 and its phosphorylation by p38. J Biol Chem. 2007; 283(7):3816-26. DOI: 10.1074/jbc.M704724200. View

14.

Ameri K, Harris A . Activating transcription factor 4. Int J Biochem Cell Biol. 2007; 40(1):14-21. DOI: 10.1016/j.biocel.2007.01.020. View

15.

Otto F, Thornell A, Crompton T, Denzel A, Gilmour K, Rosewell I . Cbfa1, a candidate gene for cleidocranial dysplasia syndrome, is essential for osteoblast differentiation and bone development. Cell. 1997; 89(5):765-71. DOI: 10.1016/s0092-8674(00)80259-7. View

16.

Ducy P, Zhang R, Geoffroy V, Ridall A, Karsenty G . Osf2/Cbfa1: a transcriptional activator of osteoblast differentiation. Cell. 1997; 89(5):747-54. DOI: 10.1016/s0092-8674(00)80257-3. View

17.

Fei Y, Xiao L, Hurley M . Fibroblast growth factor 2 positively regulates expression of activating transcription factor 4 in osteoblasts. Biochem Biophys Res Commun. 2009; 391(1):335-9. PMC: 2930756. DOI: 10.1016/j.bbrc.2009.11.059. View

18.

Ritchie H, Hou H, Veis A, Butler W . Cloning and sequence determination of rat dentin sialoprotein, a novel dentin protein. J Biol Chem. 1994; 269(5):3698-702. View

19.

Wu L, Yuan G, Yang G, Ortiz-Gonzalez I, Yang W, Cui Y . Immortalization and characterization of mouse floxed Bmp2/4 osteoblasts. Biochem Biophys Res Commun. 2009; 386(1):89-95. PMC: 2745638. DOI: 10.1016/j.bbrc.2009.05.144. View

20.

Yuan G, Wang Y, Gluhak-Heinrich J, Yang G, Chen L, Li T . Tissue-specific expression of dentin sialophosphoprotein (DSPP) and its polymorphisms in mouse tissues. Cell Biol Int. 2009; 33(8):816-29. PMC: 2725224. DOI: 10.1016/j.cellbi.2009.05.001. View