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Evaluation of MCF10A as a Reliable Model for Normal Human Mammary Epithelial Cells.

Qu Y, Han B, Yu Y, Yao W, Bose S, Karlan BY, Giuliano AE, Cui X - PLoS ONE (2015)

Bottom Line: However, there is limited knowledge about whether MCF10A cells reliably represent normal human mammary cells.When grown in suspension culture, MCF10A cells showed low mammosphere-forming ability.Our results raise a question as to whether the commonly used MCF10A cell line is a suitable model for human mammary cell studies.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, California, United States of America.

ABSTRACT
Breast cancer is the most common cancer in women and a leading cause of cancer-related deaths for women worldwide. Various cell models have been developed to study breast cancer tumorigenesis, metastasis, and drug sensitivity. The MCF10A human mammary epithelial cell line is a widely used in vitro model for studying normal breast cell function and transformation. However, there is limited knowledge about whether MCF10A cells reliably represent normal human mammary cells. MCF10A cells were grown in monolayer, suspension (mammosphere culture), three-dimensional (3D) "on-top" Matrigel, 3D "cell-embedded" Matrigel, or mixed Matrigel/collagen I gel. Suspension culture was performed with the MammoCult medium and low-attachment culture plates. Cells grown in 3D culture were fixed and subjected to either immunofluorescence staining or embedding and sectioning followed by immunohistochemistry and immunofluorescence staining. Cells or slides were stained for protein markers commonly used to identify mammary progenitor and epithelial cells. MCF10A cells expressed markers representing luminal, basal, and progenitor phenotypes in two-dimensional (2D) culture. When grown in suspension culture, MCF10A cells showed low mammosphere-forming ability. Cells in mammospheres and 3D culture expressed both luminal and basal markers. Surprisingly, the acinar structure formed by MCF10A cells in 3D culture was positive for both basal markers and the milk proteins β-casein and α-lactalbumin. MCF10A cells exhibit a unique differentiated phenotype in 3D culture which may not exist or be rare in normal human breast tissue. Our results raise a question as to whether the commonly used MCF10A cell line is a suitable model for human mammary cell studies.

No MeSH data available.


Related in: MedlinePlus

MCF10A cells embedded in mixed Matrigel/collagen I gel.Immunohistochemistry staining of basal markers (A), luminal markers (B), and breast tissue-specific markers (C) were performed in 3D-cultured MCF10A cells. Bars: 50μm. Original magnification: ×600.
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pone.0131285.g006: MCF10A cells embedded in mixed Matrigel/collagen I gel.Immunohistochemistry staining of basal markers (A), luminal markers (B), and breast tissue-specific markers (C) were performed in 3D-cultured MCF10A cells. Bars: 50μm. Original magnification: ×600.

Mentions: When embedded in the mixed gel, MCF10A cells formed acini characterized as spherical mono-layers of cells that enclose a central lumen and branches formed by elongated cells. As expected, MCF10A cells-formed branches expressed basal markers CK14 and SMA, but not P63 (Fig 6A). They also expressed luminal markers CK18 and CK8 (Fig 6B) as well as the breast tissue-specific markers CSN2 and LALBA (Fig 6C). The spherical structures showed the same marker expression (data not shown). These observations suggest that MCF10A cells in mixed Matrigel/collagen I gel also do not possess a common marker profile reflecting those of known human mammary epithelial cell types.


Evaluation of MCF10A as a Reliable Model for Normal Human Mammary Epithelial Cells.

Qu Y, Han B, Yu Y, Yao W, Bose S, Karlan BY, Giuliano AE, Cui X - PLoS ONE (2015)

MCF10A cells embedded in mixed Matrigel/collagen I gel.Immunohistochemistry staining of basal markers (A), luminal markers (B), and breast tissue-specific markers (C) were performed in 3D-cultured MCF10A cells. Bars: 50μm. Original magnification: ×600.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493126&req=5

pone.0131285.g006: MCF10A cells embedded in mixed Matrigel/collagen I gel.Immunohistochemistry staining of basal markers (A), luminal markers (B), and breast tissue-specific markers (C) were performed in 3D-cultured MCF10A cells. Bars: 50μm. Original magnification: ×600.
Mentions: When embedded in the mixed gel, MCF10A cells formed acini characterized as spherical mono-layers of cells that enclose a central lumen and branches formed by elongated cells. As expected, MCF10A cells-formed branches expressed basal markers CK14 and SMA, but not P63 (Fig 6A). They also expressed luminal markers CK18 and CK8 (Fig 6B) as well as the breast tissue-specific markers CSN2 and LALBA (Fig 6C). The spherical structures showed the same marker expression (data not shown). These observations suggest that MCF10A cells in mixed Matrigel/collagen I gel also do not possess a common marker profile reflecting those of known human mammary epithelial cell types.

Bottom Line: However, there is limited knowledge about whether MCF10A cells reliably represent normal human mammary cells.When grown in suspension culture, MCF10A cells showed low mammosphere-forming ability.Our results raise a question as to whether the commonly used MCF10A cell line is a suitable model for human mammary cell studies.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, California, United States of America.

ABSTRACT
Breast cancer is the most common cancer in women and a leading cause of cancer-related deaths for women worldwide. Various cell models have been developed to study breast cancer tumorigenesis, metastasis, and drug sensitivity. The MCF10A human mammary epithelial cell line is a widely used in vitro model for studying normal breast cell function and transformation. However, there is limited knowledge about whether MCF10A cells reliably represent normal human mammary cells. MCF10A cells were grown in monolayer, suspension (mammosphere culture), three-dimensional (3D) "on-top" Matrigel, 3D "cell-embedded" Matrigel, or mixed Matrigel/collagen I gel. Suspension culture was performed with the MammoCult medium and low-attachment culture plates. Cells grown in 3D culture were fixed and subjected to either immunofluorescence staining or embedding and sectioning followed by immunohistochemistry and immunofluorescence staining. Cells or slides were stained for protein markers commonly used to identify mammary progenitor and epithelial cells. MCF10A cells expressed markers representing luminal, basal, and progenitor phenotypes in two-dimensional (2D) culture. When grown in suspension culture, MCF10A cells showed low mammosphere-forming ability. Cells in mammospheres and 3D culture expressed both luminal and basal markers. Surprisingly, the acinar structure formed by MCF10A cells in 3D culture was positive for both basal markers and the milk proteins β-casein and α-lactalbumin. MCF10A cells exhibit a unique differentiated phenotype in 3D culture which may not exist or be rare in normal human breast tissue. Our results raise a question as to whether the commonly used MCF10A cell line is a suitable model for human mammary cell studies.

No MeSH data available.


Related in: MedlinePlus