Isolation of Neural Crest Derived Chromaffin Progenitors from Adult Adrenal Medulla
Isolation of Neural Crest Derived Chromaffin Progenitors from Adult Adrenal Medulla
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DOI：
10.1002/stem.180
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发表时间：
2009-10-01
期刊：
影响因子：
5.2
通讯作者：
Ehrhart-Bornstein, Monika
中科院分区：
文献类型：
--
作者：
Chung, Kuei-Fang;Sicard, Flavie;Ehrhart-Bornstein, Monika
肾上腺髓质的嗜铬细胞是交感肾上腺谱系的神经嵴衍生细胞。与密切相关的交感神经元不同，即使在成人中也存在增殖能力细胞的亚群。在这里，我们描述了分离，扩增，并在体外表征增殖能力的祖细胞从牛肾上腺髓质。与神经球类似，当阻止这些细胞粘附在培养皿上时，这些细胞生长在球体中，我们将其命名为染色球。这些色球缺乏平滑肌细胞（MYH 11）或内皮细胞（PECAM 1）的特异性mRNA。在球体形成过程中，分化的嗜铬细胞的标记物，如苯乙醇胺-N-甲基转移酶，下调，而神经祖细胞标记物巢蛋白，波形蛋白，武藏1，神经生长因子受体，以及神经嵴祖细胞的标记物，如Sox 1和Sox 9，上调。克隆分析和溴脱氧尿苷掺入分析证实了色球细胞的自我更新能力。分别使用NGF和BMP 4或地塞米松诱导的神经元或内分泌分化的分化方案。神经细胞的电生理学分析来自色球揭示了成熟神经细胞的功能特性，如河豚毒素敏感的钠通道和动作电位。我们的研究提供了证据，增殖和分化能力的嗜铬祖细胞可以从成人肾上腺髓质分离，这些细胞可能具有治疗神经退行性疾病，如帕金森氏病的潜力。干细胞2009;27：2602-2613
Chromaffin cells of the adrenal medulla are neural crest-derived 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'-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. STEM CELLS 2009;27:2602-2613