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Isolation and characterization of the primary epithelial breast cancer cells and the adjacent normal epithelial cells from Iranian women’s breast cancer tumors

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Abstract

As an experimental model, most studies rely on established human cancer cell lines; however, some genetical or phenotypical differences exist between these cells and their original tumor. Therefore, primary cells isolated directly from tissue are believed to be more biologically relevant tools for studying human and animal biology. Here, we aimed to isolate primary epithelial cancer and normal cells from breast tumors of Iranian women, for the first time. Thus, we isolated the epithelial and fibroblast cells from biopsy samples of patients with breast cancer based on differential centrifugation followed by culture in selective media. Normal epithelial cells obtained from the tissue biopsy away from the core of the tumor, based on the pathological diagnosis. Flow cytometry analysis indicated the positive immunoreactivity of the isolated epithelial cells against CD24 and Epithelial Specific Antigen (ESA/EpCAM), while they displayed a concomitant low expression of CD44 and CD49f. In contrat to fibroblasts, the qPCR data indicated the expression of luminal intracellular cytokeratin (Ck18) in both normal and cancer epithelial cells, but there was no expression of myoepithelial/basal markers, CK5 and vimentin. The epithelial cancer cells were reactive to cytokeratin 19 (CK19) antibody, whereas the normal epithelial cells were not. The expression of calmodulin-like protein (CLP) was also lower in the cancer epithelial cells than in the normal ones. In conclusion, primary epithelial normal and cancer cells, in addition to the fibroblasts were isolated and characterized from breast tumor of Iranian patients; and CLP expression is suggested as a susceptibility marker for breast cancer screening.

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Acknowledgements

This project was supported and funded by Iranian national Science Foundation (INSF) project No. 93036700. The authors are also thank full of the Research Console of Tarbiat Modares University for their support of this project. The authors are very thankful of Dr. Laleh Nikfarjam for her intellectual help during separation of the mentioned cells.

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Authors and Affiliations

  1. Department of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, P.O. Box 14115-331, Tehran, Iran

    Nassim Faridi & S. Zahra Bathaie

  2. Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran

    Saeid Abroun

  3. Iranian Biological Resource Center, Tehran, Iran

    Parvaneh Farzaneh

  4. Atieh Hospital, Tehran, Iran

    Hamid Karbasian

  5. Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, UCLA, Los Angeles, CA, USA

    Fuyuhiko Tamanoi

  6. Cancer Institute, Imam Khomeini Hospital, Tehran University of Medical Science, Tehran, Iran

    Mohammad-Ali Mohagheghi

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  1. Nassim Faridi
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Faridi, N., Bathaie, S.Z., Abroun, S. et al. Isolation and characterization of the primary epithelial breast cancer cells and the adjacent normal epithelial cells from Iranian women’s breast cancer tumors. Cytotechnology 70, 625–639 (2018). https://doi.org/10.1007/s10616-017-0159-3

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