Fip200 Stable Knockout 562 Tumor Cell Line
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Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing institution.
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| Cat. No. | T3476 |
| Name | Fip200 Stable Knockout 562 Tumor Cell Line |
| Description |
This Fip200 stable knockout cell line was generated via CRISPR/cas9 genome editing technology in 562 tumor cells - a spontaneously immortalized tumor cell line derived from tumors in Tsc1iΔEC mice (Cat. T8169). This cell line is useful for studies investigating malignant lymphangiosarcoma (LAS).This cell line, characterized by the absence of Fip200 protein and impaired autophagy, is of vascular endothelial tumor cell origin and exhibits continuous growth and proliferation in vitro. It is particularly valuable for cancer research, autophagy studies, and drug development, providing insights into the molecular mechanisms by which autophagy influences cancer cell behavior. Additionally, it serves as a crucial model for gene function analysis, especially regarding Fip200 and related pathways in vascular tumor cells, aiding in the discovery of new therapeutic targets. Parental cell line: Cat. T8169 - 562 Tumor Cells |
| Organism | Mouse (M. musculus) |
| Tissue | Vascular Endothelium |
| Donor History | Cutaneous tumors in tamoxifen-induced Tsc1f/f;Scl-Cre mice |
| Growth Properties | Adherent, Fusiform |
| 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 |
For optimal cell culture, we recommend using PriCoat™ T25 Flasks (G299) or coating your preferred vessels with Applied Cell Extracellular Matrix (G422) . Dulbecco's Modified Eagle Medium (DMEM), High Glucose (TM500) + 10% FBS(Regular*) + 1X Non-Essential Amino Acids (TM068) + 25 mM HEPES (TM058) + 0.6 USP U/ml Heparin + 1X Endothelial Cell Growth Supplement (ECGS) (TM141) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂. *Do not heat-inactivate |
| 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 | We recommend using serum-free CryoGuard™ Freezing Media (TM078) or, if serum is preferred, Cryopreservation Medium (TM024). |
| Split Ratio | 1:10 to 1:20 |
| Population Doubling Time (h) | 24 |
| Expression |
WB: Tsc1 deletion, mTORC1 and MAPK signals high. IHC: CD31, Lyve-1 and Prox1 positive, mTROC1 and MAPK signals high. |
| 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. Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing individual/institution for each order. If you have any questions regarding this, please contact us at licensing@abmgood.com. 2. 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. 3. We 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). 4. 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. 5. 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. 6. 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 Cincinnati |
| 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. T3476 |
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How should I handle live cells once I receive them? |
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Please refer to our Cell Handling and Thawing Guidelines for detailed instructions on receiving, thawing, and culturing live cells: |
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What is your warranty or return policy? |
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Our warranty and return policy is outlined in abm’s Terms and Conditions, including details on product quality, limitations, and claims. For additional questions, our Order team is happy to assist and can be reached at order@abmgood.com. |
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How many times can cells divide? |
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The number of times cells can divide depends on the cell type:
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Do I need Applied Cell Extracellular Matrix (G422) if I am using PriCoat™ flasks? |
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Please refer to the Growth Conditions section of your specific product page. This section will indicate whether Applied Cell Extracellular Matrix (G422), PriCoat™ flasks, or both are recommended for optimal cell attachment and growth, as requirements may vary by cell type. |
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Yang, F., Kalantari, S., Ruan, B., Sun, S., Bian, Z., & Guan, J. L. (2023). Autophagy inhibition prevents lymphatic malformation progression to lymphangiosarcoma by decreasing osteopontin and Stat3 signaling. Nature Communications, 14(1), 978.