Immortalized Mouse Hepatic Stellate Cells - SV40

Cat. No.
T0688
Unit
1x10⁶ cells / 1.0 ml
Price
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Cat. No. T0688
Name Immortalized Mouse Hepatic Stellate Cells - SV40
Description

The Immortalized Mouse Hepatic Stellate Cells – SV40 were derived from C57/Bl6 WT mouse via enzymatic digestion and Percoll density gradient centrifugation. The primary cells were immortalized via transfection of the pCMV Simian Virus 40 Large T antigen and hygromycin B selection. The Immortalized Mouse Hepatic Stellate Cells – SV40 are useful in studies focusing on liver fibrosis since it is the major cell type involved in the formation of scar tissue. The cells are particularly applicable in studies associated with inflammatory responses, as NF-κB and inflammatory cytokines levels have been found to change in the Immortalized Mouse Hepatic Stellate Cells upon LPS stimulation. Characterization by genotype analysis, NF-κB activation assay, cytokine ELISA assays, and 3[H]-thymidine cell proliferation assay.

Unit 1x10⁶ cells / 1.0 ml
Cell Type Immortalized Cells
Organism Mouse (M. musculus)
Tissue Liver
Donor History C57/Bl6 WT mouse
Growth Properties Adherent, polygonal
Seeding Density (cells/cm2) 20,000 - 40,000
Population Doubling Time (h) 35 - 45
Immortalization Method Transfection of pCMV Simian Virus 40 Large T antigen
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 + 1% Penicillin/Streptomycin Solution (G255), 33.0°C, 5% CO₂
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.

QC

1) Genotype analysis; 2)NF-κB Activation Assay; 3) Cytokine ELISA Assays; 4) 3[H]-thymidine Cell Proliferation Assay

Application Research Use Only.
Storage Condition Vapor phase of liquid nitrogen, or below -130°C.
BioSafety II
Disclaimer
1. 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 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.

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 ICAHN School of Medicine at Mount Sinai
Print & Download Datasheet
  • Guo, J., Loke, J., Zheng, F., Hong, F., Yea, S., Fukata, M., Tarocchi, M., Abar, O. T., Huang, H., Sninsky, J. J., & Friedman, S. L. (2009). Functional linkage of cirrhosis-predictive single nucleotide polymorphisms of Toll-like receptor 4 to hepatic stellate cell responses. Hepatology (Baltimore, Md.), 49(3), 960–968. https://doi.org/10.1002/hep.22697


    Lee, J. B., Park, J. S., Shin, Y. M., Lee, D. H., Yoon, J. K., Kim, D. H., ... & Sung, H. J. (2019). Implantable Vascularized Liver Chip for Cross‐Validation of Disease Treatment with Animal Model. Advanced Functional Materials, 1900075.

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