A Defined, Controlled Culture System for Primary Bovine Chromaffin Progenitors Reveals Novel Biomarkers and Modulators
Autor: | Triantafyllos Chavakis, Graeme Eisenhofer, Stefan R. Bornstein, M Ehrhart-Bornstein, Ian Levenfus, Steve W. Poser, Jimmy Masjkur, Nan Qin, Sven Lange, Vladimir Vukicevic, Carina Arps-Forker, Andreas Androutsellis-Theotokis |
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Rok vydání: | 2014 |
Předmět: |
Cholera Toxin
Time Factors Chromaffin Cells Cellular differentiation Basic fibroblast growth factor Cell Culture Techniques Biology Angiopoietin-2 chemistry.chemical_compound Catecholamines Neural Stem Cells medicine Animals Progenitor cell Protein Kinase Inhibitors Cells Cultured Cell Proliferation Janus Kinases Cell growth Intracellular Signaling Peptides and Proteins Gene Expression Regulation Developmental Membrane Proteins Cell Differentiation Cell Biology General Medicine Tissue-Specific Progenitor and Stem Cells Molecular biology Neural stem cell medicine.anatomical_structure chemistry Adrenal Medulla Cell culture Catecholamine Cattle Fibroblast Growth Factor 2 Adrenal medulla Biomarkers Signal Transduction Transcription Factors Developmental Biology medicine.drug |
Zdroj: | Stem Cells Translational Medicine. 3:801-808 |
ISSN: | 2157-6580 2157-6564 |
DOI: | 10.5966/sctm.2013-0211 |
Popis: | We present a method to efficiently culture primary chromaffin progenitors from the adult bovine adrenal medulla in a defined, serum-free monolayer system. Tissue is dissociated and plated for expansion under support by the mitogen basic fibroblast growth factor (bFGF). The cultures, although not homogenous, contain a subpopulation of cells expressing the neural stem cell marker Hes3 that also propagate. In addition, Hes3 is also expressed in the adult adrenal medulla from where the tissue is taken. Differentiation is induced by bFGF withdrawal and switching to Neurobasal medium containing B27. Following differentiation, Hes3 expression is lost, and cells acquire morphologies and biomarker expression patterns of chromaffin cells and dopaminergic neurons. We tested the effect of different treatments that we previously showed regulate Hes3 expression and cell number in cultures of fetal and adult rodent neural stem cells. Treatment of the cultures with a combination of Delta4, Angiopoietin2, and a Janus kinase inhibitor increases cell number during the expansion phase without significantly affecting catecholamine content levels. Treatment with cholera toxin does not significantly affect cell number but reduces the ratio of epinephrine to norepinephrine content and increases the dopamine content relative to total catecholamines. These data suggest that this defined culture system can be used for target identification in drug discovery programs and that the transcription factor Hes3 may serve as a new biomarker of putative adrenomedullary chromaffin progenitor cells. |
Databáze: | OpenAIRE |
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