Dual TGFβ/BMP Pathway Inhibition Enables Expansion and Characterization of Multiple Epithelial Cell Types of the Normal and Cancerous Breast.
5'-Nucleotidase
/ genetics
Animals
Antigens, Neoplasm
/ genetics
Breast
/ metabolism
Breast Neoplasms
/ genetics
Cell Differentiation
Cell Lineage
/ genetics
Epithelial Cell Adhesion Molecule
/ genetics
Epithelial Cells
/ metabolism
Female
Flow Cytometry
Gene Expression Regulation, Neoplastic
Humans
Integrin alpha6
/ genetics
Mice
Neprilysin
/ genetics
Transforming Growth Factor beta
/ genetics
Xenograft Model Antitumor Assays
Journal
Molecular cancer research : MCR
ISSN: 1557-3125
Titre abrégé: Mol Cancer Res
Pays: United States
ID NLM: 101150042
Informations de publication
Date de publication:
07 2019
07 2019
Historique:
received:
08
02
2019
revised:
18
03
2019
accepted:
12
04
2019
pubmed:
18
4
2019
medline:
2
5
2020
entrez:
18
4
2019
Statut:
ppublish
Résumé
Functional modeling of normal breast epithelial hierarchy and stromal-epithelial cell interactions have been difficult due to inability to obtain sufficient stem-progenitor-mature epithelial and stromal cells. Recently reported epithelial reprogramming assay has partially overcome this limitation, but cross-contamination of cells from the feeder layer is a concern. The purpose of this study was to develop a feeder-layer-independent and inexpensive method to propagate multiple cell types from limited tissue resources. Cells obtained after enzymatic digestion of tissues collected at surgery or by core-needle biopsies were plated on tissue culture dishes precoated with laminin-5-rich-conditioned media from the rat bladder tumor cell line 804G and a defined growth media with inhibitors of ROCK, TGFβ, and BMP signaling. Cells were characterized by flow cytometry, mammosphere assay, 3D cultures, and xenograft studies. Cells from the healthy breasts included CD10
Identifiants
pubmed: 30992305
pii: 1541-7786.MCR-19-0165
doi: 10.1158/1541-7786.MCR-19-0165
pmc: PMC6610652
mid: NIHMS1527404
doi:
Substances chimiques
Antigens, Neoplasm
0
Epithelial Cell Adhesion Molecule
0
Integrin alpha6
0
Transforming Growth Factor beta
0
5'-Nucleotidase
EC 3.1.3.5
Neprilysin
EC 3.4.24.11
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
1556-1570Subventions
Organisme : NCI NIH HHS
ID : R03 CA195250
Pays : United States
Informations de copyright
©2019 American Association for Cancer Research.
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