| Field | Specification |
|---|---|
| Species |
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.
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Tumorigenic: No
- cultureMedium: DMEM:Ham's F12 (1:1), w: 3.1 g/L Glucose, w: 2.5 mM L-Glutamine, w: 15 mM HEPES, w: 0.5 mM Sodium pyruvate, w: 1.2 g/L NaHCO3 (Cytion article number 820400a)
- supplements: Supplement the medium with 5% horse serum, 20 ng/mL EGF, 0,5 microgram/mL Hydrocortison, 10 microgram/mL Insulin. Add 100 ng/mL cholera toxin if needed.
- dissociationReagent: Accutase
- subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
- fluidRenewal: 2 to 3 times per week
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
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