HCT116-GFP

HCT116-GFP is a genetically modified derivative of the human colorectal carcinoma cell line HCT116, engineered to constitutively express green fluorescent protein (GFP). The parental HCT116 line originates from a colon carcinoma of an adult patient and is widely used as a model of mismatch repair–deficient (MMR-deficient) colorectal cancer. Cytogenetic analyses have shown that HCT116 cells typically exhibit a near-diploid karyotype with defined chromosomal alterations, including structural rearrangements and copy number variations characteristic of microsatellite instability–positive colorectal tumors. Introduction of the GFP reporter gene enables real-time visualization of cellular behavior without substantially altering the intrinsic genetic and phenotypic properties of the parental line.

The GFP labeling in HCT116-GFP cells is commonly achieved through stable transfection, resulting in uniform fluorescence that facilitates live-cell imaging, cell tracking, and quantitative analysis of proliferation, migration, and tumor growth dynamics. This modification is particularly valuable in in vitro assays and in vivo xenograft models, where GFP expression allows non-invasive monitoring of tumor progression, metastatic dissemination, and response to therapeutic interventions. Fluorescence-based phenotypic profiling approaches have demonstrated that colon cancer cell lines, including HCT116, exhibit distinct intracellular signatures that can be captured through imaging-based methodologies, supporting the utility of fluorescent reporters such as GFP for high-content screening applications.

HCT116-GFP serves as a robust tool for investigating colorectal cancer biology, enabling detailed analysis of tumor cell behavior, microenvironmental interactions, and therapeutic efficacy in both basic and translational research contexts.

Genetic modification: Stably modified by replication-incompetent lentiviral transduction to express the ZsGreen1 green fluorescent protein reporter; maintained as a polyclonal population under puromycin selection (1–5 µg/mL). S1/BSL-1 containment.

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 200 x g for 5 minutes, carefully discard the supernatant containing freezing medium.

7. Follow the procedure described under Post-Thaw Recovery

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.