Label-free isolation and cultivation of patient-matched human mammary epithelial and stromal cells from normal breast tissue

Breast cancer is primarily derived from mammary epithelial cells, the main cell type in human mammary glands. The majority of knowledge gained thus far around breast cancer has come from research using immortalized epithelial cell lines. The use of primary cells derived from breast tissue can be use...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Maria K. Koch, Berline Murekatete, Dietmar W. Hutmacher, Larisa M. Haupt, Laura J. Bray
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	ex vivo cell isolation normal breast primary cell culture mammary epithelial cells mammary fibroblasts Cytology QH573-671
Acceso en línea:	https://doaj.org/article/14002b5d2fd5458dbb90d08eeae57cef
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:14002b5d2fd5458dbb90d08eeae57cef
record_format	dspace
spelling	oai:doaj.org-article:14002b5d2fd5458dbb90d08eeae57cef2021-11-26T04:23:48ZLabel-free isolation and cultivation of patient-matched human mammary epithelial and stromal cells from normal breast tissue0171-933510.1016/j.ejcb.2021.151187https://doaj.org/article/14002b5d2fd5458dbb90d08eeae57cef2021-09-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S0171933521000388https://doaj.org/toc/0171-9335Breast cancer is primarily derived from mammary epithelial cells, the main cell type in human mammary glands. The majority of knowledge gained thus far around breast cancer has come from research using immortalized epithelial cell lines. The use of primary cells derived from breast tissue can be used in research to provide more biological relevance representative of the heterogeneous nature of breast cancer development and metastasis in its natural microenvironment. However, the successful isolation and propagation of human primary mammary gland cells can be costly and difficult due to their complex in vivo microenvironment and sensitivity when isolated. Here, we present a gentle isolation method for viable human mammary epithelial cells (hMECs) and donor-matched human mammary fibroblasts (hMFbs) from human mammary gland tissue. We isolated, expanded and passaged the hMECs and hMFbs in vitro and characterized cultures using cell-specific markers. A total of four primary cell lines were isolated and established from normal breast tissue and characterized through various markers, including pan cytokeratin (panCK), CK14, CD44, CD31, fibronectin and vimentin by immunofluorescence. To determine functional potential for subsequent studies, epithelial cells were examined via Matrigel® assays to assess spheroid development. Both cell type cultures expressed lineage specific markers with hMECs but not hMFbs forming spheroid structures in 3D Matrigel® assays. Our analyses confirm the successful isolation of two different cell phenotypes from normal breast tissues. This robust technique provides an inexpensive and accessible approach for mammary cell isolation.Maria K. KochBerline MurekateteDietmar W. HutmacherLarisa M. HauptLaura J. BrayElsevierarticleex vivo cell isolationnormal breastprimary cell culturemammary epithelial cellsmammary fibroblastsCytologyQH573-671ENEuropean Journal of Cell Biology, Vol 100, Iss 7, Pp 151187- (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	ex vivo cell isolation normal breast primary cell culture mammary epithelial cells mammary fibroblasts Cytology QH573-671
spellingShingle	ex vivo cell isolation normal breast primary cell culture mammary epithelial cells mammary fibroblasts Cytology QH573-671 Maria K. Koch Berline Murekatete Dietmar W. Hutmacher Larisa M. Haupt Laura J. Bray Label-free isolation and cultivation of patient-matched human mammary epithelial and stromal cells from normal breast tissue
description	Breast cancer is primarily derived from mammary epithelial cells, the main cell type in human mammary glands. The majority of knowledge gained thus far around breast cancer has come from research using immortalized epithelial cell lines. The use of primary cells derived from breast tissue can be used in research to provide more biological relevance representative of the heterogeneous nature of breast cancer development and metastasis in its natural microenvironment. However, the successful isolation and propagation of human primary mammary gland cells can be costly and difficult due to their complex in vivo microenvironment and sensitivity when isolated. Here, we present a gentle isolation method for viable human mammary epithelial cells (hMECs) and donor-matched human mammary fibroblasts (hMFbs) from human mammary gland tissue. We isolated, expanded and passaged the hMECs and hMFbs in vitro and characterized cultures using cell-specific markers. A total of four primary cell lines were isolated and established from normal breast tissue and characterized through various markers, including pan cytokeratin (panCK), CK14, CD44, CD31, fibronectin and vimentin by immunofluorescence. To determine functional potential for subsequent studies, epithelial cells were examined via Matrigel® assays to assess spheroid development. Both cell type cultures expressed lineage specific markers with hMECs but not hMFbs forming spheroid structures in 3D Matrigel® assays. Our analyses confirm the successful isolation of two different cell phenotypes from normal breast tissues. This robust technique provides an inexpensive and accessible approach for mammary cell isolation.
format	article
author	Maria K. Koch Berline Murekatete Dietmar W. Hutmacher Larisa M. Haupt Laura J. Bray
author_facet	Maria K. Koch Berline Murekatete Dietmar W. Hutmacher Larisa M. Haupt Laura J. Bray
author_sort	Maria K. Koch
title	Label-free isolation and cultivation of patient-matched human mammary epithelial and stromal cells from normal breast tissue
title_short	Label-free isolation and cultivation of patient-matched human mammary epithelial and stromal cells from normal breast tissue
title_full	Label-free isolation and cultivation of patient-matched human mammary epithelial and stromal cells from normal breast tissue
title_fullStr	Label-free isolation and cultivation of patient-matched human mammary epithelial and stromal cells from normal breast tissue
title_full_unstemmed	Label-free isolation and cultivation of patient-matched human mammary epithelial and stromal cells from normal breast tissue
title_sort	label-free isolation and cultivation of patient-matched human mammary epithelial and stromal cells from normal breast tissue
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/14002b5d2fd5458dbb90d08eeae57cef
work_keys_str_mv	AT mariakkoch labelfreeisolationandcultivationofpatientmatchedhumanmammaryepithelialandstromalcellsfromnormalbreasttissue AT berlinemurekatete labelfreeisolationandcultivationofpatientmatchedhumanmammaryepithelialandstromalcellsfromnormalbreasttissue AT dietmarwhutmacher labelfreeisolationandcultivationofpatientmatchedhumanmammaryepithelialandstromalcellsfromnormalbreasttissue AT larisamhaupt labelfreeisolationandcultivationofpatientmatchedhumanmammaryepithelialandstromalcellsfromnormalbreasttissue AT laurajbray labelfreeisolationandcultivationofpatientmatchedhumanmammaryepithelialandstromalcellsfromnormalbreasttissue
_version_	1718409885406724096

Label-free isolation and cultivation of patient-matched human mammary epithelial and stromal cells from normal breast tissue

Ejemplares similares