Immortalized-(Conditionally)-Mouse-Osteocytic-Cells-(IDG-SW3)-

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产品名称: Immortalized-(Conditionally)-Mouse-Osteocytic-Cells-(IDG-SW3)-

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产品展商: ABM

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Immortalized-(Conditionally)-Mouse-Osteocytic-Cells-(IDG-SW3)-

Immortalized-(Conditionally)-Mouse-Osteocytic-Cells-(IDG-SW3)- 的详细介绍

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BioSafety Level	II
Organism	3-month old Immortomouse+/- crossed with Dmp1-GFP+/- transgenic mouse
Source Organ	Long bones
Growth Properties	Adherent
Morphology	Polygonal
Recommended Seeding Density	Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.
Markers	ALP (osteoblast), E11/gp38, Dmp1 and Phex (osteocyte), SOST/sclerostin and FGF23 (late osteocyte)
Applications	For Research Use Only
Immortalization Method	Isolated from long bones of transgenic mice (in which the Dmp1 promoter drives the expression of GFP).
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).
Procedure Overview
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Propagation	Use of PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells in ECM-coated culture vessels with the following conditions. The basal medium for this cell line is Prigrow III medium available in abm, Cat. No. TM003. To make the complete growth medium, add the following components to the basal medium: 10% fetal bovine serum (TM999), 50U/ml IFN γ and Penicillin/Streptomycin (G255). Atmosphere: air, 95%; Carbon dioxide (CO2), 5%; Temperature: 33.0°C. To induce osteogenesis, replace the culture medium with the following osteogenic media: Prigrow III medium (TM003) with 10% fetal bovine serum (TM999), 50 µg/ml ascorbic acid, 4 mM β-glycerophosphate and Penicillin/Streptomycin (G255) in the absence of IFN γ at 37.0°C. Change the media every three days. GFP expression should be observed by day 4 under differentiation conditions, and become strong by day 10-15 of culture.
Preservation	.
Quality Control	(1) Presence and absence of SV40 large T-antigen, under permissive and non-permissive conditions, were confirmed using western blot. (2) Fluorescent imaging was used to measure Dmp1 promoter driven GFP expression in osteogenic conditions. (3) Von Kossa staining was used to show focal nodular mineralization and Alizarin red S staining was used to detect calcium deposition of the differentiated cells.
Disclaimer	1. The CoA for this product (provided upon request) verifies the cell-type specific gene expression via RT-PCR only. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. 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 strongly recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. 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. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping). 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 application(s). Please contact a technical service representative for more information. 4. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and provided for informational purposes only. abm does not warrant that such information has been shown to be accurate.

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