Characterization of immortalized human mammary epithelial cell line HMEC 2.6

Primary human mammary epithelial cells have a limited life span which makes it difficult to study them in vitro for most purposes. To overcome this problem, we have developed a cell line that was immortalized using defined genetic elements, and we have characterized this immortalized non-tumorigenic...

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Main Authors: Pooja S Joshi, Vishnu Modur, JiMing Cheng, Kathy Robinson, Krishna Rao
Format: Article
Language:English
Published: IOS Press 2017-10-01
Series:Tumor Biology
Online Access:https://doi.org/10.1177/1010428317724283
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spelling doaj-64b5f681c6ef4168ad00913179097dd32021-05-02T18:27:30ZengIOS PressTumor Biology1423-03802017-10-013910.1177/1010428317724283Characterization of immortalized human mammary epithelial cell line HMEC 2.6Pooja S Joshi0Vishnu Modur1JiMing Cheng2Kathy Robinson3Krishna Rao4Department of Medical Microbiology, Immunology and Cell Biology, Southern Illinois University School of Medicine, Springfield, IL, USADepartment of Pediatrics and Cincinnati Children’s Hospital, University of Cincinnati, Cincinnati, OH, USAFor You Dentistry, 477 Union Ave., Bridgewater, NJSimmons Cancer Institute at Southern Illinois University, Springfield, IL, USASimmons Cancer Institute at Southern Illinois University, Springfield, IL, USAPrimary human mammary epithelial cells have a limited life span which makes it difficult to study them in vitro for most purposes. To overcome this problem, we have developed a cell line that was immortalized using defined genetic elements, and we have characterized this immortalized non-tumorigenic human mammary epithelial cell line to establish it as a potential model system. human mammary epithelial cells were obtained from a healthy individual undergoing reduction mammoplasty at SIU School of Medicine. The cells were transduced with CDK4R24C followed by transduction with human telomerase reverse transcriptase. Post all manipulation, the cells displayed a normal cell cycle phase distribution and were near diploid in nature, which was confirmed by flow cytometry and karyotyping. In vitro studies showed that the cells were anchorage dependent and were non-invasive in nature. The cell line expressed basal epithelial markers such as cytokeratin 7, CD10, and p63 and was negative for the expression of estrogen receptor and progesterone receptor. Upon G-band karyotyping, the cell line displayed the presence of a few cytogenic abnormalities, including trisomy 20 and trisomy 7, which are also commonly present in other immortalized mammary cell lines. Furthermore, the benign nature of these cells was confirmed by multiple in vitro and in vivo experiments. Therefore, we think that this cell line could serve as a good model to understand the molecular mechanisms involved in the development and progression of breast cancer and to also assess the effect of novel therapeutics on human mammary epithelial cells.https://doi.org/10.1177/1010428317724283
collection DOAJ
language English
format Article
sources DOAJ
author Pooja S Joshi
Vishnu Modur
JiMing Cheng
Kathy Robinson
Krishna Rao
spellingShingle Pooja S Joshi
Vishnu Modur
JiMing Cheng
Kathy Robinson
Krishna Rao
Characterization of immortalized human mammary epithelial cell line HMEC 2.6
Tumor Biology
author_facet Pooja S Joshi
Vishnu Modur
JiMing Cheng
Kathy Robinson
Krishna Rao
author_sort Pooja S Joshi
title Characterization of immortalized human mammary epithelial cell line HMEC 2.6
title_short Characterization of immortalized human mammary epithelial cell line HMEC 2.6
title_full Characterization of immortalized human mammary epithelial cell line HMEC 2.6
title_fullStr Characterization of immortalized human mammary epithelial cell line HMEC 2.6
title_full_unstemmed Characterization of immortalized human mammary epithelial cell line HMEC 2.6
title_sort characterization of immortalized human mammary epithelial cell line hmec 2.6
publisher IOS Press
series Tumor Biology
issn 1423-0380
publishDate 2017-10-01
description Primary human mammary epithelial cells have a limited life span which makes it difficult to study them in vitro for most purposes. To overcome this problem, we have developed a cell line that was immortalized using defined genetic elements, and we have characterized this immortalized non-tumorigenic human mammary epithelial cell line to establish it as a potential model system. human mammary epithelial cells were obtained from a healthy individual undergoing reduction mammoplasty at SIU School of Medicine. The cells were transduced with CDK4R24C followed by transduction with human telomerase reverse transcriptase. Post all manipulation, the cells displayed a normal cell cycle phase distribution and were near diploid in nature, which was confirmed by flow cytometry and karyotyping. In vitro studies showed that the cells were anchorage dependent and were non-invasive in nature. The cell line expressed basal epithelial markers such as cytokeratin 7, CD10, and p63 and was negative for the expression of estrogen receptor and progesterone receptor. Upon G-band karyotyping, the cell line displayed the presence of a few cytogenic abnormalities, including trisomy 20 and trisomy 7, which are also commonly present in other immortalized mammary cell lines. Furthermore, the benign nature of these cells was confirmed by multiple in vitro and in vivo experiments. Therefore, we think that this cell line could serve as a good model to understand the molecular mechanisms involved in the development and progression of breast cancer and to also assess the effect of novel therapeutics on human mammary epithelial cells.
url https://doi.org/10.1177/1010428317724283
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