Three-dimensional culture models of normal and malignant breast epithelial cells

Genee Y. Lee, Paraic A. Kenny, Eva H. Lee, Mina J. Bissell

Research output: Contribution to journalArticle

771 Citations (Scopus)

Abstract

Extracellular matrix is a key regulator of normal homeostasis and tissue phenotype. Important signals are lost when cells are cultured ex vivo on two-dimensional plastic substrata. Many of these crucial microenvironmental cues may be restored using three-dimensional (3D) cultures of laminin-rich extracellular matrix (lrECM). These 3D culture assays allow phenotypic discrimination between nonmalignant and malignant mammary cells, as the former grown in a 3D context form polarized, growth-arrested acinus-like colonies whereas the latter form disorganized, proliferative and nonpolar colonies. Signaling pathways that function in parallel in cells cultured on plastic become reciprocally integrated when the cells are exposed to basement membrane-like gels. Appropriate 3D culture thus provides a more physiologically relevant approach to the analysis of gene function and cell phenotype ex vivo. We describe here a robust and generalized method for the culturing of various human breast cell lines in three dimensions and describe the preparation of cellular extracts from these cultures for molecular analyses. The procedure below describes the 3D 'embedded' assay, in which cells are cultured embedded in an lrECM gel (Fig. 1). By lrECM, we refer to the solubilized extract derived from the Engelbreth-Holm-Swarm mouse sarcoma cells. For a discussion of user options regarding 3D matrices, see Box 1. Alternatively, the 3D 'on-top' assay, in which cells are cultured on top of a thin lrECM gel overlaid with a dilute solution of lrECM, may be used as described in Box 2 (Fig. 1 and Fig. 2).

Original languageEnglish (US)
Pages (from-to)359-365
Number of pages7
JournalNature Methods
Volume4
Issue number4
DOIs
StatePublished - Apr 2007
Externally publishedYes

Fingerprint

Laminin
Extracellular Matrix
Breast
Epithelial Cells
Cultured Cells
Gels
Plastics
Experimental Sarcomas
Phenotype
Basement Membrane
Cues
Homeostasis
Cell Line
Growth
Genes

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Biology
  • Cell Biology

Cite this

Three-dimensional culture models of normal and malignant breast epithelial cells. / Lee, Genee Y.; Kenny, Paraic A.; Lee, Eva H.; Bissell, Mina J.

In: Nature Methods, Vol. 4, No. 4, 04.2007, p. 359-365.

Research output: Contribution to journalArticle

Lee, Genee Y. ; Kenny, Paraic A. ; Lee, Eva H. ; Bissell, Mina J. / Three-dimensional culture models of normal and malignant breast epithelial cells. In: Nature Methods. 2007 ; Vol. 4, No. 4. pp. 359-365.
@article{5387d4c70af945f9a77deb2634b94423,
title = "Three-dimensional culture models of normal and malignant breast epithelial cells",
abstract = "Extracellular matrix is a key regulator of normal homeostasis and tissue phenotype. Important signals are lost when cells are cultured ex vivo on two-dimensional plastic substrata. Many of these crucial microenvironmental cues may be restored using three-dimensional (3D) cultures of laminin-rich extracellular matrix (lrECM). These 3D culture assays allow phenotypic discrimination between nonmalignant and malignant mammary cells, as the former grown in a 3D context form polarized, growth-arrested acinus-like colonies whereas the latter form disorganized, proliferative and nonpolar colonies. Signaling pathways that function in parallel in cells cultured on plastic become reciprocally integrated when the cells are exposed to basement membrane-like gels. Appropriate 3D culture thus provides a more physiologically relevant approach to the analysis of gene function and cell phenotype ex vivo. We describe here a robust and generalized method for the culturing of various human breast cell lines in three dimensions and describe the preparation of cellular extracts from these cultures for molecular analyses. The procedure below describes the 3D 'embedded' assay, in which cells are cultured embedded in an lrECM gel (Fig. 1). By lrECM, we refer to the solubilized extract derived from the Engelbreth-Holm-Swarm mouse sarcoma cells. For a discussion of user options regarding 3D matrices, see Box 1. Alternatively, the 3D 'on-top' assay, in which cells are cultured on top of a thin lrECM gel overlaid with a dilute solution of lrECM, may be used as described in Box 2 (Fig. 1 and Fig. 2).",
author = "Lee, {Genee Y.} and Kenny, {Paraic A.} and Lee, {Eva H.} and Bissell, {Mina J.}",
year = "2007",
month = "4",
doi = "10.1038/nmeth1015",
language = "English (US)",
volume = "4",
pages = "359--365",
journal = "Nature Methods",
issn = "1548-7091",
publisher = "Nature Publishing Group",
number = "4",

}

TY - JOUR

T1 - Three-dimensional culture models of normal and malignant breast epithelial cells

AU - Lee, Genee Y.

AU - Kenny, Paraic A.

AU - Lee, Eva H.

AU - Bissell, Mina J.

PY - 2007/4

Y1 - 2007/4

N2 - Extracellular matrix is a key regulator of normal homeostasis and tissue phenotype. Important signals are lost when cells are cultured ex vivo on two-dimensional plastic substrata. Many of these crucial microenvironmental cues may be restored using three-dimensional (3D) cultures of laminin-rich extracellular matrix (lrECM). These 3D culture assays allow phenotypic discrimination between nonmalignant and malignant mammary cells, as the former grown in a 3D context form polarized, growth-arrested acinus-like colonies whereas the latter form disorganized, proliferative and nonpolar colonies. Signaling pathways that function in parallel in cells cultured on plastic become reciprocally integrated when the cells are exposed to basement membrane-like gels. Appropriate 3D culture thus provides a more physiologically relevant approach to the analysis of gene function and cell phenotype ex vivo. We describe here a robust and generalized method for the culturing of various human breast cell lines in three dimensions and describe the preparation of cellular extracts from these cultures for molecular analyses. The procedure below describes the 3D 'embedded' assay, in which cells are cultured embedded in an lrECM gel (Fig. 1). By lrECM, we refer to the solubilized extract derived from the Engelbreth-Holm-Swarm mouse sarcoma cells. For a discussion of user options regarding 3D matrices, see Box 1. Alternatively, the 3D 'on-top' assay, in which cells are cultured on top of a thin lrECM gel overlaid with a dilute solution of lrECM, may be used as described in Box 2 (Fig. 1 and Fig. 2).

AB - Extracellular matrix is a key regulator of normal homeostasis and tissue phenotype. Important signals are lost when cells are cultured ex vivo on two-dimensional plastic substrata. Many of these crucial microenvironmental cues may be restored using three-dimensional (3D) cultures of laminin-rich extracellular matrix (lrECM). These 3D culture assays allow phenotypic discrimination between nonmalignant and malignant mammary cells, as the former grown in a 3D context form polarized, growth-arrested acinus-like colonies whereas the latter form disorganized, proliferative and nonpolar colonies. Signaling pathways that function in parallel in cells cultured on plastic become reciprocally integrated when the cells are exposed to basement membrane-like gels. Appropriate 3D culture thus provides a more physiologically relevant approach to the analysis of gene function and cell phenotype ex vivo. We describe here a robust and generalized method for the culturing of various human breast cell lines in three dimensions and describe the preparation of cellular extracts from these cultures for molecular analyses. The procedure below describes the 3D 'embedded' assay, in which cells are cultured embedded in an lrECM gel (Fig. 1). By lrECM, we refer to the solubilized extract derived from the Engelbreth-Holm-Swarm mouse sarcoma cells. For a discussion of user options regarding 3D matrices, see Box 1. Alternatively, the 3D 'on-top' assay, in which cells are cultured on top of a thin lrECM gel overlaid with a dilute solution of lrECM, may be used as described in Box 2 (Fig. 1 and Fig. 2).

UR - http://www.scopus.com/inward/record.url?scp=34247864012&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34247864012&partnerID=8YFLogxK

U2 - 10.1038/nmeth1015

DO - 10.1038/nmeth1015

M3 - Article

C2 - 17396127

AN - SCOPUS:34247864012

VL - 4

SP - 359

EP - 365

JO - Nature Methods

JF - Nature Methods

SN - 1548-7091

IS - 4

ER -