MDA-MB-231-GFP

MDA-MB-231-GFP is a fluorescently labeled variant of the widely used MDA-MB-231 human breast cancer cell line, engineered to express green fluorescent protein (GFP) via lentiviral transduction. This modification enables real-time visualization and quantification of tumor cell dynamics both in vitro and in vivo, facilitating detailed analysis of tumor-stroma interactions, cellular proliferation, and metastatic behavior. The parental MDA-MB-231 line originates from a pleural effusion of a patient with triple-negative breast cancer (TNBC) and exhibits aggressive, invasive behavior with a mesenchymal phenotype, making it a cornerstone model for studying TNBC pathophysiology and treatment resistance.

In co-culture experiments with human mesenchymal stem/stromal cells (MSCs), MDA-MB-231-GFP cells have demonstrated significantly enhanced proliferation and tumor-promoting behavior. Studies showed that direct contact with MSCs, rather than soluble factors alone, is critical for this effect. Specifically, co-culture with MSCs led to a 39.5% increase in MDA-MB-231-GFP cell proliferation after four days compared to monoculture, and induced expression of CD90 on a subset of breast cancer cells—a marker not expressed under standard conditions. This MSC-induced CD90 expression required direct cell-cell interaction and was partially inhibited by blocking gap junctions or Notch signaling, indicating the involvement of specific intercellular communication pathways.

In vivo, co-injection of MDA-MB-231-GFP cells with MSCs into immunodeficient NOD/scid mice resulted in approximately tenfold increased tumor volume and enhanced metastatic potential compared to injection of cancer cells alone. These tumors exhibited elevated vascularization and higher viability, and retained a minority CD90-positive population, reinforcing the in vitro findings. Together, these studies position MDA-MB-231-GFP as a robust model for investigating tumor-stroma interactions, MSC-induced phenotypic plasticity, and mechanisms of tumor progression in triple-negative breast cancer.

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.