Immortalized Mouse Osteocytic Cells - Conditionally (IDG-SW3)

Cat. No.
T0231
Unit
1x10⁶ cells / 1.0 ml
Price
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Cat. No. T0231
Name Immortalized Mouse Osteocytic Cells - Conditionally (IDG-SW3)
Description

IDG-SW3 represent a non-homogenous population progressing from early osteoblasts to late osteocytic. These cells express functional SV40 large T antigen that is induced in the presence of IFN γ under permissive temperature (33°C) and express GFP under control of the dentin matrix (Dmp1) promoter. Initial culture shows osteoblastic phenotype, however when under mineralizing conditions, the cells start to express early osteocyte markers such as E11/podoplanin, followed by Dmp1 and mature markers SOST and FGF23 by 21-28 days of culture.

Similar to osteocytes in vivo, these cells respond to hormonal signals such as PTH and 1,25-dihydroxyvitamin-D3. IDG-SW3 cells can be maintained in both 2D and 3D cultures and are useful for study of osteocytes at various differentiation stages. It should be noted that these cultures may contain a mixture of osteoblasts and osteocytes at different stages of differentiation and for studies requiring enriched osteocytes, the Dmp1-GFP cells should be isolated by flow cytometry (FACS).The presence and absence of SV40 large T-antigen under permissive and non-permissive conditions, were confirmed using western blot.

Organism Mouse (M. musculus)
Tissue Bone
Donor History 3-month old Immortomouse+/- crossed with Dmp1-GFP+/- transgenic mouse
Growth Properties Adherent, polygonal
Cell Type Immortalized Cells
Unit 1x10⁶ cells / 1.0 ml
Storage Condition Vapor phase of liquid nitrogen, or below -130°C.
Shipping Conditions Ship with dry ice.
Product Format Frozen
Intended Use This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
BioSafety II
Certificate of Analysis For batch-specific test results, refer to the applicable certificate of analysis that can be found at www.abmgood.com.
Growth Conditions Use of PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. PriGrow III (TM003) + 10% FBS + 50 ng/ml rmIFNG (Z200085) + 1% Penicillin/Streptomycin Solution (G255), 33.0°C, 5% CO₂
Unpacking and Storage Instructions

1. Visually examine the packaging containers for signs of leakage or breakage.

2. Immediately transfer frozen cells from dry ice packaging to a temperature below -130°C, preferably in liquid nitrogen vapor phase storage, until ready for use.

To ensure the highest level of viability, thaw the vial and initiate culture as soon as possible upon receipt. If continued storage is desired, the vial should only be stored below -130°C or in liquid nitrogen vapor phase. Do not store at -70°C, as it will result in loss of viability.


Thawing Protocol

1. Thaw cells quickly in a 37°C water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination.

2. Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions.

3. Transfer the cell suspension into a 15ml sterile conical tube containing 5ml of pre-warmed, complete growth media. Centrifuge cells at 125xg for 5-7 minutes.

4. Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in the recommended pre-warmed, complete growth media and dispense into a T25 culture flask.

5. Incubate the cells at the recommended conditions.

Subculture Protocol

Volumes given below are for a T75 flask; proportionally increase or decrease the volume as required per culture vessel size. Subculture cells once the culture vessel is 80% confluent.

1. Aspirate the culture media, and add 2-3ml of pre-warmed 0.25% Trypsin-EDTA to the culture vessel.

2. Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes). Cells that are difficult to detach can be put in 37°C, for several minutes to facilitate detachment.

3. Neutralize Trypsin-EDTA by adding an equal volume of the complete growth media into the culture vessel.

4. Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 125xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type.

5. Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. Add appropriate aliquots of the cell suspension to new culture vessels, as desired.

6. Incubate the cells at the recommended conditions.

Cryopreservation Cryopreservation Medium (TM024), or complete growth media with 10% DMSO.
Seeding Density (cells/cm2) 25,000 - 30,000
Population Doubling Time (h) 40 - 50
Immortalization Method Isolated from long bones of transgenic mice (in which the Dmp1 promoter drives the expression of GFP)
Expression ALP (osteoblast), E11/gp38, Dmp1 and Phex (osteocyte), SOST/sclerostin and FGF23 (late osteocyte)
Material Citation If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. T0231.
Warranty abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period”.
Disclaimer
1. All test parameters provided in the CoA are conducted using abm's standardized culture system and The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 μg, Cat.# C207, $450.00) or cell lysate (100 μg, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final.

2. We recommend live cell shipments for ease of cell transfer and this option can be requested at the time of order placement. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location.

3. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic or any other non-RUO application(s).

4. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature are provided for informational purposes only. abm does not warrant that such information has been shown to be accurate.

5. abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period".
Depositor University of Missouri
Application Research Use Only.
Material Citation If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. T0231
Print & Download Datasheet
  • Woo, S. M., Rosser, J., Dusevich, V., Kalajzic, I., & Bonewald, L. F. (2011). Cell line IDG-SW3 replicates osteoblast-to-late-osteocyte differentiation in vitro and accelerates bone formation in vivo. Journal of Bone and Mineral Research, 26(11), 2634–2646. https://doi.org/10.1002/jbmr.465

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