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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 in different culture systems.(A) MCF10A cells in monolayer 2D culture. (B) MCF10A cells were cultured in ultra-low attachment 96-well plates with the MammoCult medium. (C) MCF10A cells in “on-top” of Matrigel. (D) MCF10A cells embedded in Matrigel. (E) MCF10A cells embedded in mixed Matrigel/collagen I gel. Spheres and branches in 3D culture were indicated by black arrow heads and black arrows, respectively. Bars: 100μm. Original magnification: ×200.
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pone.0131285.g001: MCF10A cells in different culture systems.(A) MCF10A cells in monolayer 2D culture. (B) MCF10A cells were cultured in ultra-low attachment 96-well plates with the MammoCult medium. (C) MCF10A cells in “on-top” of Matrigel. (D) MCF10A cells embedded in Matrigel. (E) MCF10A cells embedded in mixed Matrigel/collagen I gel. Spheres and branches in 3D culture were indicated by black arrow heads and black arrows, respectively. Bars: 100μm. Original magnification: ×200.

Mentions: We first examined the morphologies of MCF10A cells in different culture systems. Regular 2D culture of MCF10A cells yielded a cuboidal epithelial morphology (Fig 1A). When using suspension culture to enrich the stem/progenitor cell populations, MCF10A cells formed mammospheres (Fig 1B). In 3D culture of “on-top” Matrigel, MCF10A cells produced spheroid structures (Fig 1C), as previously discovered [14]. When embedded in Matrigel, the majority of MCF10A cells also formed spheroid structures (Fig 1D). We also tested MCF10A cell growth in mixed Matrigel/collagen I gel. Collagen I has been used in the in vitro 3D culture system because its mechanical tension on cells facilitates their 3D structure formation. In addition, the mixture of Matrigel and collagen I was found to create a better microenvironment for breast epithelial cells to grow branches as well as acini [13]. As expected, spheroid (Fig 1E, black arrow heads) and branch structures (Fig 1E, black arrows) were observed when the cells were embedded in mixed Matrigel/collagen I gel. These data suggest that MCF10A cells can form distinct structures in different culture conditions.


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 in different culture systems.(A) MCF10A cells in monolayer 2D culture. (B) MCF10A cells were cultured in ultra-low attachment 96-well plates with the MammoCult medium. (C) MCF10A cells in “on-top” of Matrigel. (D) MCF10A cells embedded in Matrigel. (E) MCF10A cells embedded in mixed Matrigel/collagen I gel. Spheres and branches in 3D culture were indicated by black arrow heads and black arrows, respectively. Bars: 100μm. Original magnification: ×200.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0131285.g001: MCF10A cells in different culture systems.(A) MCF10A cells in monolayer 2D culture. (B) MCF10A cells were cultured in ultra-low attachment 96-well plates with the MammoCult medium. (C) MCF10A cells in “on-top” of Matrigel. (D) MCF10A cells embedded in Matrigel. (E) MCF10A cells embedded in mixed Matrigel/collagen I gel. Spheres and branches in 3D culture were indicated by black arrow heads and black arrows, respectively. Bars: 100μm. Original magnification: ×200.
Mentions: We first examined the morphologies of MCF10A cells in different culture systems. Regular 2D culture of MCF10A cells yielded a cuboidal epithelial morphology (Fig 1A). When using suspension culture to enrich the stem/progenitor cell populations, MCF10A cells formed mammospheres (Fig 1B). In 3D culture of “on-top” Matrigel, MCF10A cells produced spheroid structures (Fig 1C), as previously discovered [14]. When embedded in Matrigel, the majority of MCF10A cells also formed spheroid structures (Fig 1D). We also tested MCF10A cell growth in mixed Matrigel/collagen I gel. Collagen I has been used in the in vitro 3D culture system because its mechanical tension on cells facilitates their 3D structure formation. In addition, the mixture of Matrigel and collagen I was found to create a better microenvironment for breast epithelial cells to grow branches as well as acini [13]. As expected, spheroid (Fig 1E, black arrow heads) and branch structures (Fig 1E, black arrows) were observed when the cells were embedded in mixed Matrigel/collagen I gel. These data suggest that MCF10A cells can form distinct structures in different culture conditions.

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