MDA-kb2

The MDA-kb2 cell line is a human breast cancer cell line derived from an adult patient. These cells are estrogen receptor (ER) negative and androgen receptor (AR) positive, making them valuable for studies involving androgen signaling pathways and their implications in breast cancer. The MDA-kb2 cell line was derived from the breast cancer cell line, MDA-MB-453, by stable transfection with a mouse mammary tumor virus (MMTV)-Luc-neo reporter gene construct. This genetic modification allows for the use of MDA-kb2 cells in bioassays for androgenic and anti-androgenic activities, where they are often utilized in-Luc reporter assays due to their stable transfection with a-Luc reporter gene under the control of an androgen-responsive promoter.

Due to their specific receptor profile, MDA-kb2 cells provide a crucial model for investigating the role of androgens in breast cancer progression and for testing the efficacy of potential therapeutic agents targeting AR pathways. These cells are cultured in Leibovitz's L-15 medium supplemented with 10% fetal bovine serum, under conditions that do not require CO₂ supplementation, which is an atypical characteristic compared to many other cell lines. The unique properties of MDA-kb2 cells make them an indispensable tool in both basic research and pharmaceutical development, particularly in understanding hormone receptor interactions in breast 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.