Cementocyte-Like Cell Line (IDG-CM6)
Clonal cementocyte-like cell line, IDG-CM6 replicates cementoblast to late cementocyte differentiation.
Highlights:
- Can be expanded under permissive conditions, at 33°C in the presence of IFN-γ, and then allowed to resume their original phenotype, at 37°C in the absence of IFN-γ
- Dmp1-GFP-negative and T-antigen-positive under immortalizing conditions, but Dmp1-GFP-positive and T-antigen-negative under osteogenic conditions
- Like osteoblasts, they express alkaline phosphatase and produce and mineralize a type I collagen matrix
- Like early osteocytes, they express E11/gp38/podoplanin and Dmp-1
- Like late osteocytes, they develop a dendritic morphology and express SOST/sclerostin
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.
Clonal cementocyte-like cell line, IDG-CM6 replicates cementoblast to late cementocyte differentiation.
Highlights:
- Can be expanded under permissive conditions, at 33°C in the presence of IFN-γ, and then allowed to resume their original phenotype, at 37°C in the absence of IFN-γ
- Dmp1-GFP-negative and T-antigen-positive under immortalizing conditions, but Dmp1-GFP-positive and T-antigen-negative under osteogenic conditions
- Like osteoblasts, they express alkaline phosphatase and produce and mineralize a type I collagen matrix
- Like early osteocytes, they express E11/gp38/podoplanin and Dmp-1
- Like late osteocytes, they develop a dendritic morphology and express SOST/sclerostin
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.
| Catalog Number | Product | DataSheet | Size | AVAILABILITY | Price | Qty |
|---|
| Product Type: | Cell Line |
| Name: | IDG-CM6 |
| Cell Type: | Late cementoblast/cementocyte |
| Organism: | Mouse |
| 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. |
| Species: | Mouse |
| 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. |
| Growth Conditions: |
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 |
| Shipped: | Dry Ice |
Experimental Protocol:
- For proliferation/expansion: incubate at 33°C
- For differentiation: incubate at 37°C
- Plate the cells at a density of 4x10^4 cells/cm2, in proliferation medium and incubate at 33°C, 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 usually observed at 3-4 days of culture under differentiation conditions, and should be strong by days 10-14 of culture.
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.
- Zhao N, Nociti Jr FH, Duan P, Prideaux M, Zhao H, Foster BL, Somerman MJ, Bonewald LF. Isolation and Functional Analysis of an Immortalized Murine Cementocyte Cell Line, IDG-CM6. J Bone Miner Res. 2016 Feb: 31(2): 430-442. DOI: 10.1002/jbmr.2690
- Zhao N, Foster BL, Bonewald LF. The Cementocyte-An Osteocyte Relative? J Dental Res. 2016, Vol 95 (7) 734-741. DOI: 10.1177/0022034516641898
- Woo SM, Rosser J, Dusevich V, Kalajzic I, Bonewald LF. Cell line IDG-SW3 replicates osteoblast-to-late-osteocyte differentiation in vitro and accelerates bone formation in vivo. J Bone Miner Res. 2011 Nov; 26(11):2634-46. DOI: 10.1002/jbmr.465
If you publish research with this product, please let us know so we can cite your paper.
