Clonal cementocyte-like cell line, IDG-CM6 replicates cementoblast to late cementocyte differentiation.
The clonal cementocyte-like cell line IDG-CM6 (for Immortomouse/Dmp1-GFP-CM6), was derived from the apical portion of 1st to 3rd mandibular molar tooth roots from 3 mo. old female mice carrying a DMP-1 (Dentin matrix protein-1) promoter driving GFP (green fluorescent protein) crossed with the Immortomouse, which expresses a thermolabile SV40 large Tumor-antigen, regulated by IFN- γ (interferon-gamma). This cell line should be useful to study cementoblast to cementocyte transition, mechanisms for biomineralization cementocyte function and regulation of SOST/sclerostin and DMP-1.
From the laboratory of Lynda F. Bonewald, PhD, University of Missouri - Kansas City.
|Product Type:||Cell Line|
|Cell Type:||Late cementoblast/cementocyte|
|Morphology:||Adherent, similar to an osteoblast/cementoblast-like cell which, under osteogenic conditions, can form a mineralized matrix, differentiate into an cementocyte-like cell, and express GFP driven by the Dentin Matrix Protein 1 promoter.|
|Source:||Apical portion of tooth roots from 1st to 3rd mandibular molars|
|Biosafety Level:||BSL 1|
|Subculturing:||Under permissive conditions at 33C, when at 80-90% confluent, use a 0.05% Trypsin/ 0.53mM EDTA solution to detach cells. Split 1:5 to 1:10 based on your need. Cells grow best when cultured at higher densities; at low densities, proliferation can be slow. Exchange media every 2-3 days.|
Proliferation medium: AlphaMEM (containing L-glutamine and deoxyribonucleosides); supplemented with 10% FBS; penicillin-streptomycin at 100U/ml-100ug/ml; Recombinant Mouse Interferon-gamma (INF-g) range 8-50 U/ml; for growing at 33°C.
Differentiation medium: AlphaMEM (L-glutamine and deoxyribonucleosides); supplemented with 10% Fetal Bovine Serum; penicillin-streptomycin at 100U/ml-100ug/ml; approximately 50µg/ml Ascorbic Acid and 4mM β-glycerophosphate; for growing at 37°C.
Grown on dishes coated with [0.15 mg/ml] rat tail type I collagen.
|Cryopreservation:||60% AlphaMEM, 30% FBS, 10% DMSO, at 1-2 x 10^6 cells/vial/1ml|
|Comments:||For experimental purposes, plate the cells at a density of 4x10^4 cells/cm2, in proliferation medium and incubate at 33C, 5% CO2 , until confluent, called Day 0. At Day 0, switch to differentiation medium, and incubate at 37°C, 5% CO2, exchanging media every 2-3 days. The GFP expression, controlled by the DMP1 promoter, is mildly observed at 5-7 days of culture under differentiation conditions, and usually peaks around day 35 of culture|
|Storage:||After slowly freezing cryovials in a -80C freezer overnight, transfer to a liquid nitrogen tank for long term storage|
NOTE: Mineralization and gene expression may be Fetal Bovine Serum dependent; testing and optimization of different serum lots/batches may be necessary. Mineralization and gene expression may be CO2 dependent; testing and optimization of 5-10% CO2 may be necessary. Testing with our current serum showed that differentiation of the IDG-SW3 in 8% CO2 resulted in an increase of mineral, and in SOST and DMP1 gene expression, when compared to cells differentiated in 5% CO2.
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