| Field | Specification |
|---|---|
| Product Format | Frozen |
| Product Type | |
| Shipping | |
| Storage |
Overview
HCT116 infected with lentivirus carrying pWPI-GFP encoding constitutively active (CA)-MEK5 (MEK5DD; Ser313 and Thr317 activating phosphorylation residues substituted with Asp). Control cell line is available DN-MEK5 Stably Expressing HCT116 Cell Line (T6445).
Key elements and design rationale
- Model identity: CA-MEK5 Stably Expressing HCT116 Cell Line is supplied as an engineered cell line derived from Human colon.
- Growth properties: Adherent, epithelial-like
- Growth conditions: PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) are recommended for optimal cell culture. McCoy's 5A Medium (TM506) + 10% FBS(Regular*) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂. *Do not heat-inactivate
- Engineering / immortalization: Gfp reporter expression.
- Product format: Frozen, BSL-2
This cell-based model is generally used in cancer biology, phenotype comparison, and response profiling studies. Donor/background information is available for contextual interpretation.
Biological background
This model supports studies in cancer biology, phenotype comparison, and response profiling. It can be used to examine morphology, growth behavior, and experimental responses in cultured cells. Donor/background information provided for this product: Male, 48, Caucasian, Colorectal carcinoma.
Research relevance and current trends
- Engineered cell lines are widely used for reporter-based readouts, perturbation studies, and assay optimization in reproducible culture systems.
- Reporter or transgene-bearing models are often compared with matched parental or control cells to interpret signal changes in context.
- Expression trends are typically evaluated alongside passage number, selection pressure, and baseline growth behavior.
Common research applications
- Cancer biology studies that compare proliferation-associated behavior, morphology, and pathway responses in vitro.
- Assay development for treatment response, reporter monitoring, or phenotype comparison under matched culture conditions.
- Side-by-side comparison of engineered versus parental background characteristics when relevant to the study design.
Changes in morphology, growth rate, viability, or reporter signal are typically interpreted together with passage history, culture matrix, and the specified growth conditions for the model.
Notes for experimental interpretation
- Morphology, doubling behavior, and reporter or marker output can shift with passage number, substrate choice, and medium composition; these variables should be recorded alongside experimental readouts.
- Matched controls such as parental cells, untreated cultures, or parallel cultures maintained under identical conditions help distinguish background effects from biology of interest.
Culture and product details
- Growth Conditions: PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) are recommended for optimal cell culture. McCoy's 5A Medium (TM506) + 10% FBS(Regular*) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂. *Do not heat-inactivate
- Seeding Density (cells/cm²): 10,000
- 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.
- 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.
- 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.
- 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.
- Incubate the cells at the recommended conditions.
- Aspirate the culture media, and add 2-3ml of pre-warmed 0.25% Trypsin-EDTA to the culture vessel.
- 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.
- Neutralize Trypsin-EDTA by adding an equal volume of the complete growth media into the culture vessel.
- 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.
- 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.
- Incubate the cells at the recommended conditions.
How should I handle live cells once I receive them?
https://www.abmgood.com/immortalized-cells-documents.html
Following these guidelines will help ensure optimal cell viability and performance.
Why are these cells classified as biosafety level II?
What is your warranty or return policy?
Please refer to the following link for full information:
https://www.abmgood.com/terms
For additional questions, our Order team is happy to assist and can be reached at order@abmgood.com.
How many times can cells divide?
Primary cells have a limited lifespan and will undergo a finite number of population doublings before entering senescence. The exact number varies by cell type and culture conditions.
Immortalized cell lines are capable of extended or indefinite proliferation under proper culture conditions, although growth characteristics may vary between lines.
Do I need Applied Cell Extracellular Matrix (G422) if I am using PriCoat™ flasks?
Cell line sourcing and selection (species, tissue, and disease model matching) · Stable cell line engineering (overexpression, knockdown, knockout via CRISPR/Cas9, shRNA, sgRNA) · Reporter gene integration (GFP, RFP, luciferase, fluorescent/bioluminescent constructs) · Genome editing and knockin (point mutations, tagged endogenous proteins, conditional alleles) · Inducible expression systems (Tet-On/Off and regulatable constructs) · Drug resistance marker selection (puromycin, G418, hygromycin, and others) · Custom growth and media optimisation for specific assay requirements · Scale-up production for high-throughput screening campaigns · Authentication and QC services (STR profiling, mycoplasma testing, viability assessment). Talk to a Scientist or contact support@biohippo.com.
Pereira, D. M., Gomes, S. E., Borralho, P. M., & Rodrigues, C. (2019). MEK5/ERK5 activation regulates colon cancer stem-like cell properties. Cell death discovery, 5, 68. https://doi.org/10.1038/s41420-019-0150-1
Pereira, D. M., Simões, A. E., Gomes, S. E., Castro, R. E., Carvalho, T., Rodrigues, C. M., & Borralho, P. M. (2016). MEK5/ERK5 signaling inhibition increases colon cancer cell sensitivity to 5-fluorouracil through a p53-dependent mechanism. Oncotarget, 7(23), 34322–34340. https://doi.org/10.18632/oncotarget.9107
Simões, A. E., Pereira, D. M., Gomes, S. E., Brito, H., Carvalho, T., French, A., Castro, R. E., Steer, C. J., Thibodeau, S. N., Rodrigues, C. M., & Borralho, P. M. (2015). Aberrant MEK5/ERK5 signalling contributes to human colon cancer progression via NF-κB activation. Cell death & disease, 6(4), e1718. https://doi.org/10.1038/cddis.2015.83
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