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PMID: 7785785 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Isolation of pure populations of epithelial and myoepithelial cells from the normal human mammary gland using immunomagnetic separation with Dynabeads.

Analytical biochemistry ·Vol. 226 ·No. 1 ·1995-03-20 ·Pages 91-9

Gomm JJ, Browne PJ, Coope RC, Liu QY, Buluwela L, Coombes RC

Abstract

Monoclonal antibodies to epithelial membrane antigen and common acute lymphoblastic leukemia antigen were bound to second antibody-coated magnetic microspheres. These specific antibody/bead complexes were then used to directly isolate purified epithelial and myoepithelial cells from normal breast organoid cell preparations by magnetic separation. Near homogeneous cell populations were selected with yields of 10-20 x 10(6) epithelial and myoepithelial cells per organoid preparation. Repeated purification steps allowed almost complete depletion of myoepithelial cells for RNA studies. Tissue culture of separated cell populations in appropriate defined media further ensured purity of cell type and was unimpeded by Dynabead attachment.

MeSH Terms
Antibodies, Monoclonal/immunology Base Sequence Breast/cytology Cell Separation/methods Cells, Cultured DNA, Complementary/genetics Electrophoresis, Agar Gel Epithelial Cells Female Gene Expression/genetics Humans Immunomagnetic Separation Membrane Glycoproteins/immunology Molecular Sequence Data Mucin-1 Mucins/immunology Neprilysin/immunology
Chemicals
Antibodies, Monoclonal DNA, Complementary Membrane Glycoproteins Mucin-1 Mucins Neprilysin
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Gomm J J
Department of Medical Oncology, Charing Cross and Westminster Medical School, University of London, United Kingdom.
Browne P J
Coope R C
Liu Q Y
Buluwela L
Coombes R C
Article Info
Journal
Analytical biochemistry
Abbr.
Anal Biochem
ISSN
0003-2697
Published
1995-03-20
Pages
91-9
Language
English
Region
United States
NLM ID
0370535
Subset
IM
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