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MCF-10A is a spontaneously immortalized, non-tumorigenic human mammary epithelial cell line, catalogued by ATCC as CRL-10317. It was derived from tissue of a woman with fibrocystic breast disease, and it is widely used as a non-cancer comparator and as a model that forms gland-like acini in three-dimensional culture. This page is general reference for researchers and lab staff, not a protocol and not medical advice.
Short answer
MCF-10A (also written MCF 10A or MCF10A, the same line) is an adherent human breast epithelial line. ATCC lists it as non-tumorigenic, biosafety level 1, research use only, grown in MEBM with the MEGM kit plus cholera toxin. Its best-known use is 3D culture on reconstituted basement membrane, where it forms polarized acini-like spheroids.
Definition and origin
The MCF-10 culture was established from subcutaneous mastectomy tissue of a 36-year-old parous, premenopausal woman with fibrocystic disease. Tait, Soule and Russo (Cancer Research, 1990) report that the culture began as a mortal population, MCF-10M, and spontaneously gave rise to two immortal sublines: MCF-10A, which grows attached, and MCF-10F, which grows floating. The cells were grown for more than four years in DMEM/Ham’s F-12 medium, and the authors describe luminal ductal characteristics rather than myoepithelial ones, with epithelial identity confirmed by sialomucins and keratins.
How it is used in practice
Debnath, Muthuswamy and Brugge (Methods, 2003) describe culturing MCF-10A on a reconstituted basement membrane, where the cells form polarized, growth-arrested, acini-like spheroids that recapitulate aspects of glandular architecture. In the same paper, introducing oncogenes such as ErbB2 or cyclin D1 disrupts that architecture with distinct phenotypes, which is why the line is used to study how oncogenes change epithelial organization. ATCC lists 3D cell culture and use as a transfection host as applications of CRL-10317.
Culture specification as stated by ATCC
- Medium: MEBM with the MEGM kit (Lonza CC-3150, without GA-1000) and 100 ng/ml cholera toxin. ATCC states not to filter the complete medium.
- Conditions: 37 °C, 95% air and 5% CO2; adherent growth.
- Passaging and feeding: subculture ratio of 1:3 to 1:4; medium renewed every 2 to 3 days.
- Freezing and storage: freeze medium is complete medium with 7.5% DMSO; ATCC says to store in liquid nitrogen vapor and that storage at -70 °C will result in loss of viability.
- Biosafety: BSL 1. Research use only.
ATCC also notes that lots produced before 05/20/2020 may have used a different cryopreservative, so check the lot documentation for the vial you hold. Read the current ATCC product sheet for full handling instructions rather than relying on a summary.
Origin versus culture spec versus use
| Question | What the source says | Source |
|---|---|---|
| Where did it come from? | Fibrocystic disease tissue from a 36-year-old woman; spontaneous immortalization of MCF-10M gave MCF-10A and MCF-10F | Tait et al., 1990 |
| What medium was used originally? | DMEM/Ham’s F-12 at 1.05 mM calcium or 0.04 mM low calcium | Tait et al., 1990 |
| What medium does the repository specify? | MEBM with MEGM kit and cholera toxin | ATCC CRL-10317 |
| What is the signature assay? | 3D acini-like spheroids on basement membrane; oncogene perturbation | Debnath et al., 2003 |
| Tumorigenic? | Described as non-tumorigenic | ATCC CRL-10317 |
The medium rows differ: the founding literature used a DMEM/Ham’s F-12 base and showed that calcium strongly affects morphology, while ATCC specifies a different base and supplement kit. Neither is wrong; they describe different points in the line’s history. For reproducibility, report which one you used.
Related lines
MCF-10A is often paired with cancer-derived breast lines. For a tumor-derived comparator, see the CASRAI guide to the MCF-7 cell line. Other CASRAI cell-line references include A549 cells and MDCK cells.
Where it matters for labs and supply decisions
- Named consumables: ATCC names a specific medium kit (Lonza CC-3150, without GA-1000) and a cholera toxin supplement, so substituting a different base or kit changes the conditions the repository specifies.
- Cold storage: ATCC specifies liquid nitrogen vapor storage, so a lab needs that capacity before ordering; -70 °C storage is stated to lose viability.
- Record keeping: record the name as written by the source (MCF 10A, MCF-10A, MCF10A), the catalog number CRL-10317, the lot, the medium base and calcium level, and the passage number.
- Identity: confirm identity and monitor for drift; see cell line authentication.
Common misunderstandings
Non-tumorigenic does not mean normal
The line came from fibrocystic disease tissue and became immortal spontaneously in culture. ATCC calls it non-tumorigenic, which is a statement about tumor formation, not a claim that the cells behave like primary tissue from a healthy donor.
MCF-10A and MCF-10F are different sublines
Tait et al. describe the attached line (10A) and the floating line (10F) as separate immortal derivatives of the same culture.
Spelling variants are one line
MCF-10A, MCF 10A and MCF10A refer to the same line; use the ATCC catalog number CRL-10317 to be unambiguous.
FAQ
What does MCF10A mean?
It is a common spelling of MCF-10A, the immortalized human mammary epithelial line described above.
Is MCF-10A a cancer cell line?
ATCC lists it as non-tumorigenic. It was derived from fibrocystic disease tissue, not from a carcinoma.
What biosafety level is MCF-10A?
ATCC lists BSL 1 for CRL-10317. Your institution’s biosafety committee sets local requirements.
Why is MCF-10A used in 3D culture?
On reconstituted basement membrane it forms polarized, growth-arrested acini-like spheroids, per Debnath et al. (2003).
Sources
- ATCC, MCF 10A (CRL-10317) product page, atcc.org/products/crl-10317, read 2026-11-08.
- Tait L, Soule HD, Russo J. Ultrastructural and immunocytochemical characterization of an immortalized human breast epithelial cell line, MCF-10. Cancer Res 1990, PubMed 1697506, read 2026-11-08.
- Debnath J, Muthuswamy SK, Brugge JS. Morphogenesis and oncogenesis of MCF-10A mammary epithelial acini grown in three-dimensional basement membrane cultures. Methods 2003, PubMed 12798140, read 2026-11-08.
