COLO-320

COLO-320 is a human colorectal adenocarcinoma cell line derived from a metastatic tumor. It is extensively used in research to study colorectal cancer biology, drug resistance mechanisms, and the effects of anticancer agents. The COLO-320 cells are characterized by the overexpression of various oncogenes, such as c-myc, which is known to drive cell proliferation. This cell line is also a model for investigating the role of epigenetic modifications in cancer progression, as it exhibits frequent DNA methylation and histone acetylation abnormalities that influence the expression of key tumor suppressor genes and cell cycle regulators.

Research on COLO-320 has revealed that the methylation status of the CDKN2A (p16) promoter is a critical factor in the regulation of this tumor suppressor gene, which is responsible for inhibiting the G1/S phase transition in the cell cycle. Treatment with DNA methyltransferase inhibitors like 5-aza-2’-deoxycytidine (5-aza-dC) has been shown to demethylate the CDKN2A promoter, restoring gene expression and potentially inducing growth arrest. Furthermore, histone deacetylase inhibitors (HDACis) such as trichostatin A (TSA) can modulate histone acetylation levels, affecting the expression of other genes, including p21WAF1. However, studies have indicated that histone acetylation specifically influences p21WAF1 without significantly impacting CDKN2A-associated histones, suggesting differential epigenetic regulation.

Another area of research focuses on COLO-320's role in multidrug resistance (MDR). The cell line has been used to evaluate the efficacy of various MDR modulators. Phenothiazine derivatives, for instance, have demonstrated the ability to inhibit P-glycoprotein (P-gp) activity, a protein often overexpressed in COLO-320 cells that pumps out chemotherapeutic drugs, thereby reducing their intracellular concentration and efficacy. Studies have utilized flow cytometry with rhodamine 123 efflux assays to quantify the effectiveness of these compounds in reversing MDR, providing insights into potential strategies to overcome drug resistance in colorectal cancer.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.

To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.