Immortalized mouse floxed Bmp2 dental papilla mesenchymal cell lines preserve odontoblastic phenotype and respond to BMP2.
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.
Wu
Li-an
LA
Department of Pediatric Dentistry, Dental School, The University of Texas Health Science Center, San Antonio, Texas 78229-3900, USA.
Feng
Junsheng
J
Wang
Lynn
L
Mu
Yan-dong
YD
Baker
Andrew
A
Donly
Kevin J
KJ
Gluhak-Heinrich
Jelica
J
Harris
Stephen E
SE
MacDougall
Mary
M
Chen
Shuo
S
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics / Q000378:metabolism*; D002113:Calcification, Physiologic; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D048430:Cell Shape; D003771:Dental Papilla / Q000166:cytology* / Q000502:physiology; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000502:physiology*; D051379:Mice; D008822:Mice, Transgenic; D009804:Odontoblasts / Q000166:cytology* / Q000502:physiology*; D010641:Phenotype
D015415:Biomarkers; C525719:Bmp2 protein, mouse; D055396:Bone Morphogenetic Protein 2
Science. 1988 Dec 16;242(4885):1528-34
3201241
Genesis. 2005 Jul;42(3):203-6
15986484
PLoS Genet. 2006 Dec;2(12):e216
17194222
Arch Oral Biol. 2000 Oct;45(10):857-69
10973559
Arch Oral Biol. 2005 Feb;50(2):271-7
15721161
J Clin Invest. 2003 Sep;112(6):924-34
12975477
Eur J Oral Sci. 2006 Apr;114(2):138-46
16630306
Nucleic Acids Res. 2002 Jan 15;30(2):515-22
11788714
Am J Med Genet A. 2005 Mar 1;133A(2):138-41
15666299
J Biol Chem. 2008 Feb 15;283(7):3816-26
18056716
Int J Biochem Cell Biol. 2008;40(1):14-21
17466566
Development. 2005 Dec;132(24):5601-11
16314491
Cytokine Growth Factor Rev. 1997 Mar;8(1):11-20
9174660
Development. 1996 Oct;122(10):2977-86
8898212
Cell. 1997 May 30;89(5):765-71
9182764
Cell. 1997 May 30;89(5):747-54
9182762
Biochem Biophys Res Commun. 2010 Jan 1;391(1):335-9
19913500
J Biol Chem. 1994 Feb 4;269(5):3698-702
8106414
Biochem Biophys Res Commun. 2009 Aug 14;386(1):89-95
19501574
J Biol Chem. 2008 Jul 18;283(29):20198-208
18492670
Growth Factors. 2004 Dec;22(4):233-41
15621726
Cell. 2004 Apr 30;117(3):387-98
15109498
J Biol Chem. 2008 Oct 24;283(43):29119-25
18703512
Genes Dev. 1996 Jul 1;10(13):1580-94
8682290
Sex Dev. 2008;2(3):134-41
18769073
Int Endod J. 2008 Jul;41(7):609-16
18479370
Nat Genet. 1997 Jul;16(3):307-10
9207800
Nat Genet. 2006 Dec;38(12):1424-9
17099713
J Endod. 1998 Mar;24(3):171-5
9558581
Nucleic Acids Res. 1997 Aug 15;25(16):3326-31
9241248
Cells Tissues Organs. 2007;186(1):70-7
17627120
J Biol Chem. 2005 Aug 19;280(33):29717-27
15980071
J Biol Chem. 2005 Sep 2;280(35):30689-96
16000305
J Dent Res. 2007 May;86(5):392-9
17452557
Dev Biol. 2006 Jul 15;295(2):580-8
16730346
Hum Mol Genet. 2002 Oct 1;11(21):2559-65
12354781
Am J Med Genet A. 2008 Feb 1;146A(3):343-9
18203197
Connect Tissue Res. 1998;37(3-4):233-49
9862224
Hum Mol Genet. 1998 Mar;7(3):563-9
9467018
Eur J Oral Sci. 2007 Feb;115(1):48-56
17305716
J Cell Sci. 2003 May 1;116(Pt 9):1647-8
12665545
Kidney Int. 2000 Jun;57(6):2207-14
10844590
Mol Cell Biol. 2007 Aug;27(15):5468-78
17515606
Cytokine Growth Factor Rev. 2009 Oct-Dec;20(5-6):475-80
19892583
Arch Oral Biol. 1994 Sep;39(9):775-82
7802613
Cytokine Growth Factor Rev. 2005 Jun;16(3):369-76
15878301
Cell. 2002 Jan 11;108(1):17-29
11792318
Cell. 1997 May 30;89(5):773-9
9182765
Nat Genet. 2001 Feb;27(2):201-4
11175790
Cell. 1997 May 30;89(5):755-64
9182763
Development. 1999 Jul;126(13):2911-20
10357935
J Biol Chem. 2008 Jul 11;283(28):19359-70
18424784
Crit Rev Oral Biol Med. 1993;4(5):679-728
8292714
Am J Hum Genet. 1999 Nov;65(5):1268-78
10521292
Proc Natl Acad Sci U S A. 2007 Jan 16;104(3):1027-32
17209010
J Bone Miner Res. 1997 Dec;12(12):2040-9
9421236
Science. 1965 Nov 12;150(3698):893-9
5319761
Am J Med Genet A. 2006 Dec 1;140(23):2536-46
16955410
J Dent Res. 2009 Nov;88(11):1020-5
19828890
J Biol Chem. 2008 Mar 28;283(13):8412-22
18204048
Connect Tissue Res. 1995;33(1-3):97-103
7554969
J Oral Pathol Med. 1993 Apr;22(4):175-9
8391080