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Development of an odontoblast in vitro model to study
dentin mineralization.
David Magne, Gilles Bluteau, Serena Lopez-Cazaux, Pierre Weiss, Paul Pilet,
Helena Ritchie, Guy Daculsi, Jérôme Guicheux
To cite this version:
David Magne, Gilles Bluteau, Serena Lopez-Cazaux, Pierre Weiss, Paul Pilet, et al.. Development of an odontoblast in vitro model to study dentin mineralization.. Connective Tissue Research, Taylor & Francis, 2004, 45 (2), pp.101-8. �10.1080/03008200490464839�. �inserm-00175147�
Table 1: primers used for RT-PCR analysis (sequences, annealing temperature (T), size of
PCR products (bp), and reference, GAPDH: glyceraldehyde phosphate dehydrogenase, ALP: alkaline phosphatase, DMP-1: dentin matrix protein 1, DSPP: dentin sialophosphoprotein).
F 5’-ACC ACA GTC CAT GCC ATC AC-3’ R 5’-TCC ACC ACC CTG TTG CTG TA-3’
GAPDH
F 5’-CAC ATC CAG GAA CCG CAG CAC-3’ R 5’CCT TAC TCT CCT TTG CCT C-3’
DSPP
F 5’-GAA GAC TGT TAT CCT TAC G-3’ R 5’-GTG ATC CCC TTT AGA TTC C-3’
DMP-1
F 5’-CTT CTT GCT AGT GGA AGG-3’ R 5’-CCT GGT AGT TGT TGT GAG-3’
ALP
F 5’-CCA ATC TGG TTC CCT CCC AC-3’ R 5’-TGG TAA GGT TGA ATG CAC TT-3’
COLL-1
Forward (F) and Reverse (R) sequence
Name T annealing bp reference
62°C 51°C 62°C 62°C 60°C 452 181 672 346 330 (27) (27) (28) (29) unpublished
0 2000 4000 6000 8000 10000 12000 14000 Days in culture 2 4 7 14 21 C el ls p er w el l (x 1 0 3 )
*
* *
* * *
0 1 2 3 4 0 0,5 1 1,5 2 2,5 3 0 0,5 1 1,5 2 0 0,5 1 1,5 COLL-1 181 pb ALP 346 pb DMP1 672 pb DSPP 330 pb GAPDH 452 pb 7 9 11 16 20 -RT Days in culture C O L L -1 /G A P D H (A U ) A L P /G A P D H (A U ) D M P -1 /G A P D H (A U ) D S P P /G A P D H (A U ) 7 9 11 16 20 Days in culture
50 100 150 200 250 300 350 400 450 8 9 10 11 12 14 16 20 µ g C a /m g p ro t Days in culture
*
*
*
*
*
Pi-
+
-
+
-
+
-
+
-
+
-
+
-
+
-
+
A
B
28 4000 Wavenumbers (cm-1) 1000 1500 νννν1νννν3PO4 Amide I νννν2CO3C
D
Wavenumbers (cm-1) 1200 960 1125 1030 1020 11100 50 100 150 µ g C a /m g p ro t TGF-ββββ