Isolation, characterization, and differentiation of progenitor cells from human adult adrenal medulla

Magda M. Santana, Kuei Fang Chung, Vladimir Vukicevic, Joana Rosmaninho-Salgado, Waldemar Kanczkowski, Vera Cortez, Karl Hackmann, Carlos A. Bastos, Alfredo Mota, Evelin Schrock, Stefan R. Bornstein, Cláudia Cavadas, Monika Ehrhart-Bornstein*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

35 Citations (Scopus)

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.
Original languageEnglish
Pages (from-to)783-791
Number of pages9
JournalStem cells translational medicine
Volume1
Issue number11
DOIs
Publication statusPublished - 2012
Externally publishedYes

Keywords

  • Adult stem cells
  • Culture
  • Differentiation
  • Nestin
  • Neural differentiation
  • Progenitor cells
  • Somatic stem cells

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