A novel method for rapid enrichment of lactotrophs from dispersed anterior pituitary cells of the rat.
A polyclonal antibody to the rat D2 dopamine (DA) receptor was rapidly and covalently attached to surface-activated polystyrene cultureware (MicroCEL-Lector plates). Addition of a suspension of dispersed rat anterior pituitary cells resulted in the rapid (within 1 h) selection of cells possessing D2 DA receptors (i.e. lactotrophs). Four-fold enrichment (from about 20% in the suspension to about 80%) was routinely obtained, as judged by prolactin (PRL) immunostaining. The enriched cells were virtually free of fibroblasts and were much more homogeneous in appearance than untreated cells after 3 days in culture. Lactotroph-enriched cell cultures displayed similar functional characteristics as untreated cells when assessed by determining dose-response curves for inhibition of PRL secretion by the DA agonist N-propylnorapomorphine. This method may be generally applicable for the selective enrichment and purification of desired cell types from heterogeneous mixtures in tissue dispersions.
Meller
E
E
Millhauser Laboratories, Department of Psychiatry, New York University School of Medicine.
Bohmaker
K
K
Puza
T
T
Diamond
J
J
Lew
J Y
JY
Schütte
M
M
Okrongly
D
D
Goldstein
M
M
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D001058:Apomorphine / Q000031:analogs & derivatives / Q000494:pharmacology; D002448:Cell Adhesion; D002478:Cells, Cultured; D003584:Cytological Techniques; D015259:Dopamine Agents / Q000494:pharmacology; D008297:Male; D008969:Molecular Sequence Data; D010903:Pituitary Gland, Anterior / Q000187:drug effects / Q000378:metabolism*; D011137:Polystyrenes; D011388:Prolactin / Q000276:immunology / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D017448:Receptors, Dopamine D2 / Q000187:drug effects / Q000276:immunology
D000906:Antibodies; D015259:Dopamine Agents; D011137:Polystyrenes; D017448:Receptors, Dopamine D2; C009074:N-n-propylnorapomorphine; D011388:Prolactin; D001058:Apomorphine