title: Evaluation of MCF10A as a Reliable Model for Normal Human Mammary Epithelial Cells.
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.
authors.affiliation: Department of Surgery, Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, California, United States of America.
authors.forename: Ying
authors.initials: Y
authors.lastname: Qu
authors.affiliation: Department of Surgery, Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, California, United States of America.
authors.forename: Bingchen
authors.initials: B
authors.lastname: Han
authors.affiliation: Department of Surgery, Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, California, United States of America.
authors.forename: Yi
authors.initials: Y
authors.lastname: Yu
authors.affiliation: Department of Radiology, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, 600 Yishan Road, Shanghai, China.
authors.forename: Weiwu
authors.initials: W
authors.lastname: Yao
authors.affiliation: Department of Pathology, Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, California, United States of America.
authors.forename: Shikha
authors.initials: S
authors.lastname: Bose
authors.affiliation: Department of Obstetrics and Gynecology, Women's Cancer Program, Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, California, United States of America.
authors.forename: Beth Y
authors.initials: BY
authors.lastname: Karlan
authors.affiliation: Department of Surgery, Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, California, United States of America.
authors.forename: Armando E
authors.initials: AE
authors.lastname: Giuliano
authors.affiliation: Department of Surgery, Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, California, United States of America; Department of Obstetrics and Gynecology, Women's Cancer Program, Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, California, United States of America.
authors.forename: Xiaojiang
authors.initials: X
authors.lastname: Cui
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000032:analysis; D001940:Breast / Q000166:cytology*; D002448:Cell Adhesion; D018929:Cell Culture Techniques; D002460:Cell Line / Q000737:chemistry / Q000166:cytology* / Q000187:drug effects; D048430:Cell Shape; D003094:Collagen; D024042:Collagen Type I; D003470:Culture Media / Q000494:pharmacology; D004338:Drug Combinations; D004847:Epithelial Cells / Q000737:chemistry / Q000166:cytology* / Q000187:drug effects; D005260:Female; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D007633:Keratins / Q000032:analysis; D007797:Laminin; D008565:Membrane Proteins / Q000032:analysis; D008894:Milk Proteins / Q000032:analysis; D010641:Phenotype; D011509:Proteoglycans; D011960:Receptors, Estrogen / Q000032:analysis; D011980:Receptors, Progesterone / Q000032:analysis; D013234:Stem Cells / Q000737:chemistry / Q000166:cytology; D014746:Vimentin / Q000032:analysis
chemical_list: D015415:Biomarkers; C081876:CKAP4 protein, human; D024042:Collagen Type I; D003470:Culture Media; D004338:Drug Combinations; D036341:Intercellular Signaling Peptides and Proteins; D007797:Laminin; D008565:Membrane Proteins; D008894:Milk Proteins; D011509:Proteoglycans; D011960:Receptors, Estrogen; D011980:Receptors, Progesterone; D014746:Vimentin; C059485:matrigel; D007633:Keratins; D003094:Collagen