Isolation of Neural Crest Derived Chromaffin Progenitors from Adult Adrenal Medulla

Abstract

Chromaffin cells of the adrenal medulla are neural crestderived cells of the sympathoadrenal lineage. Unlike the closely-related sympathetic neurons, a subpopulation of proliferation-competent cells exists even in the adult. Here, we describe the isolation, expansion, and in vitro characterization of proliferation-competent progenitor cells from the bovine adrenal medulla. Similar to neurospheres, these cells, when prevented from adherence to the culture dish, grew in spheres, which we named chromospheres. These chromospheres were devoid of mRNA specific for smooth muscle cells (MYH11) or endothelial cells (PECAM1). During sphere formation, markers for differentiated chromaffin cells, such as phenylethanolamine-N-methyl transferase, were downregulated while neural progenitor markers nestin, vimentin, musashi 1, and nerve growth factor receptor, as well as markers of neural crest progenitor cells such as Sox1 and Sox9, were upregulated. Clonal analysis and bromo-2 0 -deoxyuridine-incorporation analysis demonstrated the self-renewing capacity of chromosphere cells. Differentiation protocols using NGF and BMP4 or dexamethasone induced neuronal or endocrine differentiation, respectively. Electrophysiological analyses of neural cells derived from chromospheres revealed functional properties of mature nerve cells, such as tetrodotoxin-sensitive sodium channels and action potentials. Our study provides evidence that proliferation and differentiation competent chromaffin progenitor cells can be isolated from adult adrenal medulla and that these cells might harbor the potential for the treatment of neurodegenerative diseases, such as Parkinson's disease.

Adrenal cortical and chromaffin stem cells: Is there a common progeny related to stress adaptation?

“…Since we have previously shown that human and bovine medullary progenitors express Nestin (Chung et al, 2009;Santana et al, 2012;Vukicevic et al, 2012), we were able to take advantage of the Nestin-GFP mouse model, where enhanced green fluorescent protein is expressed under the regulation of the Nes gene regulatory element (Mignone et al, 2004). In this mouse we detected Nestin-positive cells through expression of GFP, both in the adrenal medulla and cortex ( Fig.…”

“…We have for the first time been able to prove the existence of proliferation and differentiation-competent sympathoadrenal progenitor cells within the adult adrenal medulla and have established methods for their isolation from bovine (Chung et al, 2009;Ehrhart-Bornstein et al, 2010;Masjkur et al, 2014) and human (Santana et al, 2012) adrenals. Under low-attachment cell culture conditions, where adherence to the culture dish is prevented, these cells express progenitor markers, have in vitro self-renewing capacity and grow in spheres, which we called chromospheres.…”

“…5 differentiating into cells of the chromaffin and neural lineages (Chung et al, 2009;Santana et al, 2012;Vukicevic et al, 2012).…”

Transplantation of bovine adrenocortical cells encapsulated in alginate

“…BACs and chromaffin cells were isolated from bovine adrenals of freshly slaughtered 1-3-y-old cattle by collagenase digestion, as previously described (17,31,32).…”

18F-FDOPA: A Multiple-Target Molecule

“…The origin of neuroendocrine cells is local, throughout the gut wall in the gastroenteropancreatic NETs (2); the catecholamineproducing NETs arise from chromaffin cells and their common sympathoadrenal progenitors, which migrate from the neural ectoderm (6). Neuroendocrine cells store the transported and decarboxylated amines in cytoplasmic neurosecretory granules that vary in size, shape, and capacity to store peptide hormones.…”