MCF10A

The MCF10A human mammary epithelial cell line, established from the mammary gland of a 36-year-old female with fibrocystic disease, serves as a model for studying the intricacies of normal breast cell function, transformation, and the epithelial to mesenchymal transition critical in invasive breast carcinoma transition.

As a non-tumorigenic epithelial cell line derived from benign proliferative breast tissue, MCF10A cells are instrumental in mammary cell studies, offering insights into breast tumor progression and the dynamics of tumor cells in mammospheres. MCF10 A cells, characterized by their three-dimensional growth in collagen and their ability to form acinar structures in mixed Matrigel, provide a reliable model for analyzing the impact of oncogenes and studying the mammosphere formation, which is crucial for understanding the properties of mammary progenitor cells and their role in cancer research.

The MCF10A cell line, while exhibiting a basal-like phenotype, express a combination of luminal and stem-like markers, as well as epithelial-cell markers such as cytokeratins and milk proteins. Their responsiveness to insulin, glucocorticoids, cholera enterotoxin, and epidermal growth factor (EGF) underscores the importance of growth factors and hormones in the proliferation and survival of human breast tissue cells.

The MCF 10A model, provides a window into the genomic signaling pathways that govern cell behavior and phenotype in 3D culture, offering a platform for immunohistochemistry and immunofluorescence staining to visualize cellular processes.
These cells are crucial for studying the transition of mammary cells during breast cancer development, including the role of lipid oxidation product genotoxicity and the impact of dietary components like soybean trypsin inhibitor on cell function. Furthermore, the MCF 10A cell line's comparison with other lines such as MCF7 (which is tumorigenic and estrogen receptor-positive) and MCF10F (another non-tumorigenic line but with different characteristics) enriches breast cancer research by providing diverse models for understanding the spectrum of non-invasive to highly metastatic phenotypes.

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.