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Print Version |
| BioSafety Level |
II |
| Organism |
Mouse |
| Source Organ |
Adipose tissue |
| Growth Properties |
Adherent |
| Morphology |
Fibroblast-like |
| Recommended Seeding Density |
Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions. |
| Applications |
For Research Use Only |
| Immortalization Method |
Spontaneous immortalization |
| Description |
Immortalized Mouse Subcutaneous Adipose Multipotent Mesenchymal Cells (m17.ASC-GFP) were genetically modified by lentiviral vectors carrying green fluorescent protein
(GFP). When injected in the spleen of NOD/SCID-gnull monocrotaline-treated mice, m17.ASC-GFP cells efficiently engrafted in the liver, candidating this cell line as a valuable stem cell model.
It stably expresses the stem cell markers Sca-1, c-Kit/CD117, nestin, nucleostemin, CD44, and CD106, but is negative for the embryo stem cell markers Nanog, Oct4, and Sox2. The cells were endowed with multipotency and can be induced to differentiate toward osteogenic, chondrogenic, adipogenic, and cardiogenic phenotypes. It displays a normal karyotype and stable telomeres, neither proliferates when plated in soft agar nor gives rise to tumors when injected subcutaneously in NOD/SCID-g null mice. The cell line is thus recommended as a valuable tool for a number of applications in ex vivo tissue engineering to study the differentiation mechanisms involved in tissue repair, as well as a model for both pharmacological and toxicological studies.
abm also offers the Immortalized Mouse Subcutaneous Adipose Multipotent Mesenchymal Cells (m17.ASC) (abm, T0542) which does carry GFP expression.
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| 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 base medium for the cell line is Claycomb medium (Sigma-Aldrich). To make the completed growth medium, add the following components to the base medium: 2mM L-glutamine (G275), 10% of fetal bovine serum (TM999), 1% Penicillin/Streptomycin Solution (G255). Atmosphere: air: 95%, CO?: 5%; Temperature: 37.0°C. |
| Quality Control |
1) RT-PCR and Cytofluorimetry were performed on the cDNA from the cells to confirm the expression of stem cell markers Sca-1, c-kit, Islet 1, nestin, and nucleostemin.
2) Cells after differentiation were specifically stained with alizarin red, adipo-red and Alcian blue, and the tissue-specific collagen II antibodies, respectively to confirm osteogenic, adipogenic, and chondrogenic phenotypes the cells’ multipotency.
3) G-banding analysis for macroscopic chromosome alterations over time compared with normal mouse karyotypes.
4) GFP transduction was confirmed by culturing with antibiotics.
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| 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. |
