title: Isolation, characterization, and differentiation of progenitor cells from human adult adrenal medulla.
abstract: Chromaffin cells, sympathetic neurons of the dorsal ganglia, and the intermediate small intensely fluorescent cells derive from a common neural crest progenitor cell. Contrary to the closely related sympathetic nervous system, within the adult adrenal medulla a subpopulation of undifferentiated progenitor cells persists, and recently, we established a method to isolate and differentiate these progenitor cells from adult bovine adrenals. However, no studies have elucidated the existence of adrenal progenitor cells within the human adrenal medulla. Here we describe the isolation, characterization, and differentiation of chromaffin progenitor cells obtained from adult human adrenals. Human chromaffin progenitor cells were cultured in low-attachment conditions for 10-12 days as free-floating spheres in the presence of fibroblast growth factor-2 (FGF-2) and epidermal growth factor. These primary human chromosphere cultures were characterized by the expression of several progenitor markers, including nestin, CD133, Notch1, nerve growth factor receptor, Snai2, Sox9, Sox10, Phox2b, and Ascl1 on the molecular level and of Sox9 on the immunohistochemical level. In opposition, phenylethanolamine N-methyltransferase (PNMT), a marker for differentiated chromaffin cells, significantly decreased after 12 days in culture. Moreover, when plated on poly-l-lysine/laminin-coated slides in the presence of FGF-2, human chromaffin progenitor cells were able to differentiate into two distinct neuron-like cell types, tyrosine hydroxylase (TH)(+)/β-3-tubulin(+) cells and TH(-)/β-3-tubulin(+) cells, and into chromaffin cells (TH(+)/PNMT(+)). This study demonstrates the presence of progenitor cells in the human adrenal medulla and reveals their potential use in regenerative medicine, especially in the treatment of neuroendocrine and neurodegenerative diseases.
authors.affiliation: University of Coimbra, Coimbra, Portugal.
authors.forename: Magda M
authors.initials: MM
authors.lastname: Santana
authors.forename: Kuei-Fang
authors.initials: KF
authors.lastname: Chung
authors.forename: Vladimir
authors.initials: V
authors.lastname: Vukicevic
authors.forename: Joana
authors.initials: J
authors.lastname: Rosmaninho-Salgado
authors.forename: Waldemar
authors.initials: W
authors.lastname: Kanczkowski
authors.forename: Vera
authors.initials: V
authors.lastname: Cortez
authors.forename: Klaus
authors.initials: K
authors.lastname: Hackmann
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Bastos
authors.forename: Alfredo
authors.initials: A
authors.lastname: Mota
authors.forename: Evelin
authors.initials: E
authors.lastname: Schrock
authors.forename: Stefan R
authors.initials: SR
authors.lastname: Bornstein
authors.forename: Cláudia
authors.initials: C
authors.lastname: Cavadas
authors.forename: Monika
authors.initials: M
authors.lastname: Ehrhart-Bornstein
mesh_terms: D000313:Adrenal Medulla / Q000166:cytology*; D015415:Biomarkers; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival; D002478:Cells, Cultured; D019439:Chromaffin Cells / Q000378:metabolism*; D004815:Epidermal Growth Factor / Q000494:pharmacology; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D006801:Humans; D007381:Intermediate Filament Proteins / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism; D064231:Nestin; D012313:RNA / Q000032:analysis; D013234:Stem Cells / Q000166:cytology*; D014404:Tubulin / Q000378:metabolism; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
chemical_list: D015415:Biomarkers; D007381:Intermediate Filament Proteins; C577484:NES protein, human; D009419:Nerve Tissue Proteins; D064231:Nestin; C498585:TUBB3 protein, human; D014404:Tubulin; D016222:Fibroblast Growth Factor 2; D004815:Epidermal Growth Factor; D012313:RNA; D014446:Tyrosine 3-Monooxygenase