MCA-3D

The MCA-3D cell line is derived from primary mouse epidermal cultures that exhibit resistance to calcium-induced terminal differentiation. These cells were initially treated with the carcinogens N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) or 7,12-dimethylbenz[a]anthracene (DMBA), and subsequently exposed to 12-O-tetradecanoylphorbol-13-acetate (TPA). The resistance to terminal differentiation was assessed by elevating calcium levels in the culture medium to 1.2 mM, which selectively allows for the growth of transformed cells while normal cells typically undergo terminal differentiation and death.

The MCA-3D cell line displays an epithelial morphology and forms well-defined colonies in culture. Ultrastructural analysis reveals that MCA-3D cells contain keratin filaments and desmosomes, which are indicative of their epithelial origin and suggest maintenance of some degree of normal keratinocyte differentiation. However, the exact abundance of these structures can vary among subpopulations within the line.

MCA-3D cells have been tested for tumorigenicity by subcutaneous injection into syngeneic Balb/c neonates, with results indicating that this line is non-tumorigenic, even after prolonged culture in high calcium conditions. Additionally, the MCA-3D cells do not grow in soft agar, further supporting their non-malignant phenotype. Biochemical assays for gamma glutamyl transpeptidase (GGT) activity and transglutaminase activity have shown that MCA-3D cells are negative for GGT, and their transglutaminase activity does not correlate with tumorigenic potential, aligning with their non-tumorigenic classification.

Overall, the MCA-3D cell line serves as a model for studying the early stages of carcinogenesis and the factors that influence the progression from preneoplastic lesions to fully malignant tumors.

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.