Isolation of pure populations of epithelial and myoepithelial cells from the normal human mammary gland using immunomagnetic separation with Dynabeads.
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.
Gomm
J J
JJ
Department of Medical Oncology, Charing Cross and Westminster Medical School, University of London, United Kingdom.
Browne
P J
PJ
Coope
R C
RC
Liu
Q Y
QY
Buluwela
L
L
Coombes
R C
RC
D000911:Antibodies, Monoclonal / Q000276:immunology; D001483:Base Sequence; D001940:Breast / Q000166:cytology*; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D004847:Epithelial Cells; D005260:Female; D015870:Gene Expression / Q000235:genetics; D006801:Humans; D018189:Immunomagnetic Separation; D008562:Membrane Glycoproteins / Q000276:immunology; D008969:Molecular Sequence Data; D018396:Mucin-1; D009077:Mucins / Q000276:immunology; D015260:Neprilysin / Q000276:immunology
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D008562:Membrane Glycoproteins; D018396:Mucin-1; D009077:Mucins; D015260:Neprilysin