Immortalized Mouse Floxed Bmp2 Dental Papilla Mesenchymal Cell Lines Preserve Odontoblastic Phenotype and Respond to BMP2

Overview

Journal J Cell Physiol

Specialties Cell Biology
Physiology

Date 2010 May 12

PMID 20458728

Citations 13

Authors

Li-An Wu

Junsheng Feng

Lynn Wang

Yan-Dong Mu

Andrew Baker

Kevin J Donly

Jelica Gluhak-Heinrich

Stephen E Harris

Mary Macdougall

Shuo Chen

Affiliations

Soon will be listed here.

Abstract

Bone morphogenetic protein 2 (Bmp2) is essential for odontogensis and dentin mineralization. Generation of floxed Bmp2 dental mesenchymal cell lines is a valuable application for studying the effects of Bmp2 on dental mesenchymal cell differentiation and its signaling pathways during dentinogenesis. Limitation of the primary culture of dental mesenchymal cells has led to the development of cell lines that serve as good surrogate models for the study of dental mesenchymal cell differentiation into odontoblasts and mineralization. In this study, we established and characterized immortalized mouse floxed Bmp2 dental papilla mesenchymal cell lines, which were isolated from 1st mouse mandibular molars at postnatal day 1 and immortalized with pSV40 and clonally selected. These transfected cell lines were characterized by RT-PCR, immunohistochemistry, and analyzed for alkaline phosphatase activity and mineralization nodule formation. One of these immortalized cell lines, iBmp2-dp, displayed a higher proliferation rate, but retained the genotypic and phenotypic characteristics similar to primary cells as determined by expression of tooth-specific markers as well as demonstrated the ability to differentiate and form mineralized nodules. In addition, iBmp2-dp cells were inducible and responded to BMP2 stimulation. Thus, we for the first time described the establishment of an immortalized mouse floxed Bmp2 dental papilla mesenchyma cell line that might be used for studying the mechanisms of dental cell differentiation and dentin mineralization mediated by Bmp2 and other growth factor signaling pathways.

Citing Articles

Meta-analysis of the Mesenchymal Stem Cells Immortalization Protocols: A Guideline for Regenerative Medicine.

Stricker P, Oliveira N, Mogharbel B, Luhrs L, Irioda A, Abdelwahid E Curr Stem Cell Res Ther. 2024; 19(7):1009-1020.

PMID: 38221663 DOI: 10.2174/011574888X268464231016070900.

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.

Differential lncRNA/mRNA Expression Profiling and Functional Network Analyses in Bmp2 Deletion of Mouse Dental Papilla Cells.

Wang F, Tao R, Zhao L, Hao X, Zou Y, Lin Q Front Genet. 2022; 12:702540.

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Sinking Our Teeth in Getting Dental Stem Cells to Clinics for Bone Regeneration.

Shoushrah S, Transfeld J, Tonk C, Buchner D, Witzleben S, Sieber M Int J Mol Sci. 2021; 22(12).

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Establishment of Immortalized Laryngeal Epithelial Cells Transfected with Bmi1.

Tan J, Wang L, Mo T, Dai Y, Lu J, Liu X Cell Transplant. 2020; 29:963689720908198.

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References

Wozney J, Rosen V, Celeste A, Mitsock L, Whitters M, Kriz R . Novel regulators of bone formation: molecular clones and activities. Science. 1988; 242(4885):1528-34. DOI: 10.1126/science.3201241. View

Ma L, Martin J . Generation of a Bmp2 conditional null allele. Genesis. 2005; 42(3):203-6. DOI: 10.1002/gene.20132. View

Bandyopadhyay A, Tsuji K, Cox K, Harfe B, Rosen V, Tabin C . Genetic analysis of the roles of BMP2, BMP4, and BMP7 in limb patterning and skeletogenesis. PLoS Genet. 2006; 2(12):e216. PMC: 1713256. DOI: 10.1371/journal.pgen.0020216. View

Thonemann B, Schmalz G . Immortalization of bovine dental papilla cells with simian virus 40 large t antigen. Arch Oral Biol. 2000; 45(10):857-69. DOI: 10.1016/s0003-9969(00)00056-x. View

Priam F, Ronco V, Locker M, Bourd K, Bonnefoix M, Duchene T . New cellular models for tracking the odontoblast phenotype. Arch Oral Biol. 2005; 50(2):271-7. DOI: 10.1016/j.archoralbio.2004.10.007. View

Wu X, Li Y, Schneider A, Yu W, Rajendren G, Iqbal J . Impaired osteoblastic differentiation, reduced bone formation, and severe osteoporosis in noggin-overexpressing mice. J Clin Invest. 2003; 112(6):924-34. PMC: 193662. DOI: 10.1172/JCI15543. 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

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

10.

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

11.

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

12.

Ma L, Lu M, Schwartz R, Martin J . Bmp2 is essential for cardiac cushion epithelial-mesenchymal transition and myocardial patterning. Development. 2005; 132(24):5601-11. DOI: 10.1242/dev.02156. View

13.

Reddi A . Bone morphogenetic proteins: an unconventional approach to isolation of first mammalian morphogens. Cytokine Growth Factor Rev. 1997; 8(1):11-20. DOI: 10.1016/s1359-6101(96)00049-4. View

14.

Zhang H, Bradley A . Mice deficient for BMP2 are nonviable and have defects in amnion/chorion and cardiac development. Development. 1996; 122(10):2977-86. DOI: 10.1242/dev.122.10.2977. 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.

Duverger O, Lee D, Hassan M, Chen S, Jaisser F, Lian J . Molecular consequences of a frameshifted DLX3 mutant leading to Tricho-Dento-Osseous syndrome. J Biol Chem. 2008; 283(29):20198-208. PMC: 2459267. DOI: 10.1074/jbc.M709562200. View