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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

Immunofluorescence staining of the stem-like cell populations in MCF10A cells in 2D culture.MCF10A cells grown in monolayer were fixed and stained using the antibodies against reported mammary stem/progenitor (A) and stem-like (B) markers. Secondary antibodies were either AF488 (green)—or AF594 (red)—conjugated. DAPI was used for nuclear staining. Bars: 50μm. Original magnification: ×200.
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pone.0131285.g003: Immunofluorescence staining of the stem-like cell populations in MCF10A cells in 2D culture.MCF10A cells grown in monolayer were fixed and stained using the antibodies against reported mammary stem/progenitor (A) and stem-like (B) markers. Secondary antibodies were either AF488 (green)—or AF594 (red)—conjugated. DAPI was used for nuclear staining. Bars: 50μm. Original magnification: ×200.

Mentions: We next asked whether MCF10A cells express reported breast stem/progenitor cell markers. It was demonstrated that EpCAM+/Muc1− epithelial cells within the luminal epithelial lineage may function as precursor cells of the terminal duct lobular units in the human breast [28]. Studies also showed that breast stem/progenitor cells may possess the EpCAM+/CD49f+ [8, 29], Aldehyde dehydrogenase (ALDH1) high [30], or CD44+/CD24- [31] phenotype. In particular, ALDH activity is widely used to identify stem/progenitor cells [30], and this activity can be attributed to ALDH1A3 in stem cells [32]. Using immunofluorescence staining, we found that a high percentage of MCF10A cells exhibited the EpCAM+/Muc1-, ALDH1A3+/CD49f+, or CD44+/CD24- phenotype (Fig 3A and S3 Table). Next, we examined the expression of the stem cell-associated transcription factors Sox-2, Nanog, and Oct4 [33] in MCF10A cells. As presented in Fig 3B, MCF10A cells did not express Nanog but expressed high levels of Oct4 and Sox2. These data suggest that MCF10A cells contain sub-populations expressing stem/progenitor markers.


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)

Immunofluorescence staining of the stem-like cell populations in MCF10A cells in 2D culture.MCF10A cells grown in monolayer were fixed and stained using the antibodies against reported mammary stem/progenitor (A) and stem-like (B) markers. Secondary antibodies were either AF488 (green)—or AF594 (red)—conjugated. DAPI was used for nuclear staining. Bars: 50μm. Original magnification: ×200.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4493126&req=5

pone.0131285.g003: Immunofluorescence staining of the stem-like cell populations in MCF10A cells in 2D culture.MCF10A cells grown in monolayer were fixed and stained using the antibodies against reported mammary stem/progenitor (A) and stem-like (B) markers. Secondary antibodies were either AF488 (green)—or AF594 (red)—conjugated. DAPI was used for nuclear staining. Bars: 50μm. Original magnification: ×200.
Mentions: We next asked whether MCF10A cells express reported breast stem/progenitor cell markers. It was demonstrated that EpCAM+/Muc1− epithelial cells within the luminal epithelial lineage may function as precursor cells of the terminal duct lobular units in the human breast [28]. Studies also showed that breast stem/progenitor cells may possess the EpCAM+/CD49f+ [8, 29], Aldehyde dehydrogenase (ALDH1) high [30], or CD44+/CD24- [31] phenotype. In particular, ALDH activity is widely used to identify stem/progenitor cells [30], and this activity can be attributed to ALDH1A3 in stem cells [32]. Using immunofluorescence staining, we found that a high percentage of MCF10A cells exhibited the EpCAM+/Muc1-, ALDH1A3+/CD49f+, or CD44+/CD24- phenotype (Fig 3A and S3 Table). Next, we examined the expression of the stem cell-associated transcription factors Sox-2, Nanog, and Oct4 [33] in MCF10A cells. As presented in Fig 3B, MCF10A cells did not express Nanog but expressed high levels of Oct4 and Sox2. These data suggest that MCF10A cells contain sub-populations expressing stem/progenitor markers.

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