title: Isolation of human adrenocortical aldosterone-producing cells by a novel immunomagnetic beads method.
abstract: We detected intense CD56 immunostaining in the zona glomerulosa (ZG) and medulla of the normal human adrenal gland and therefore identified CD56, the neural cell adhesion molecule, as a membrane antigen specific for the ZG, aldosterone-producing adenoma (APA), and chromaffin cells. The APA and pheochromocytoma cells, which are histogenetically derived from the ZG and medulla, respectively, also showed intense CD56 immunostaining. Based on these findings we developed a strategy for isolating cells from the ZG and APA using CD56 immunobinding to magnetic beads. Morphology, gene expression studies, and aldosterone measurement confirmed that CD56 positive (+) cells were ZG and APA cells. Analysis of CD56+ cells under light and phase contrast microscopy evidenced that these cells formed clumps, as the ZG cells usually do; with electron microscopy they showed multiple features typical of a steroidogenic phenotype. Expression levels of the CD56 and the aldosterone synthase (CYP11B2) gene were markedly higher in CD56+ cells than CD56- cells (+1600 and +2100% increase, respectively). Moreover, aldosterone secretion was higher (+1380%) from CD56+ cells than from CD56- cells. Hence, this novel methodology allows isolation of a pure population of ZG and APA cells exhibiting multiple characteristics of the aldosterone-producing cells.
authors.affiliation: Department of Clinical and Experimental Medicine, University of Padua, 35126 Padua, Italy.
authors.forename: Brasilina
authors.initials: B
authors.lastname: Caroccia
authors.forename: Ambrogio
authors.initials: A
authors.lastname: Fassina
authors.forename: Teresa Maria
authors.initials: TM
authors.lastname: Seccia
authors.forename: Chiara
authors.initials: C
authors.lastname: Recarti
authors.forename: Lucia
authors.initials: L
authors.lastname: Petrelli
authors.forename: Anna Sandra
authors.initials: AS
authors.lastname: Belloni
authors.forename: Maria Rosa
authors.initials: MR
authors.lastname: Pelizzo
authors.forename: Gian Paolo
authors.initials: GP
authors.lastname: Rossi
mesh_terms: D000306:Adrenal Cortex Neoplasms / Q000378:metabolism*; D018246:Adrenocortical Adenoma / Q000378:metabolism*; D000450:Aldosterone / Q000378:metabolism*; D019002:CD56 Antigen / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D018189:Immunomagnetic Separation; D015384:Zona Glomerulosa / Q000166:cytology* / Q000378:metabolism
chemical_list: D019002:CD56 Antigen; D000450:Aldosterone