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.

Catalog Number Product Size AVAILABILITY Price Qty
EKC005
Cementocyte-Like Cell Line (IDG-CM6), 1 vial (Non-Profit)
1 vial In stock
Price: $699.00
EKC006
Cementocyte-Like Cell Line (IDG-CM6), 1 vial (For-Profit)
1 vial In stock
Price: $1,120.00
Specifications
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
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 ß-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
Source: Apical portion of tooth roots from 1st to 3rd mandibular molars
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
Documentation
Provider
From the laboratory of Lynda F. Bonewald, PhD, University of Missouri - Kansas City.
Comments

Experimental Protocol:

  1. For proliferation/expansion: incubate at 33°C
  2. For differentiation: incubate at 37°C
  3. 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.
  4. At Day 0, switch to differentiation medium, and incubate at 37°C, 5% CO2, exchanging media every 2-3 days.
  5. 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.

References
  1. 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
  2. Zhao N, Foster BL, Bonewald LF. The Cementocyte-An Osteocyte Relative? J Dental Res. 2016, Vol 95 (7) 734-741. DOI: 10.1177/0022034516641898
  3. 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.

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