Generation and properties of a new human ventral mesencephalic neural stem cell line

被引:43
作者
Villa, Ana [1 ]
Liste, Isabel [1 ]
Courtois, Elise T. [1 ]
Seiz, Emma G. [1 ]
Ramos, Milagros [1 ]
Meyer, Morten [2 ]
Juliusson, Bengt [3 ]
Kusk, Philip [3 ]
Martinez-Serrano, Alberto [1 ]
机构
[1] UAM, CSIC, Ctr Biol Mol Severo Ochoa, Madrid 28049, Spain
[2] Univ So Denmark, Inst Med Biol, Dept Anat & Neurobiol, DK-500 Odense C, Denmark
[3] NsGene AS, Ballerup, Denmark
基金
英国医学研究理事会;
关键词
Human neural stem cells; Parkinson's disease; Dopaminergic neurons; Ventral mesencephalon; Immortalization; FETAL NIGRAL TRANSPLANTATION; BCL-X-L; IN-VITRO; DOPAMINERGIC-NEURONS; PARKINSONS-DISEASE; PROGENITOR CELLS; NEURODEGENERATIVE DISORDERS; TYROSINE-HYDROXYLASE; GRAFT-SURVIVAL; DIFFERENTIATION;
D O I
10.1016/j.yexcr.2009.03.011
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Neural stem cells (NSCs) are powerful research tools for the design and discovery of new approaches to cell therapy in neurodegenerative diseases like Parkinson's disease. Several epigenetic and genetic strategies have been tested for long-term maintenance and expansion of these cells in vitro. Here we report the generation of a new stable cell line of human neural stem cells derived from ventral mesencephalon (hVM1) based on nu-myc immortalization. The cells expressed neural stem cell and radial glia markers like nestin, vimentin and 3CB2 under proliferation conditions. After withdrawal of Growth factors, proliferation and expression of nu-myc were dramatically reduced and the cells differentiated into astrocytes, oligodendrocytes and neurons. hVM1 cells yield a large number of dopaminergic neurons (about 12% of total cells are TH+) after differentiation, which also produce dopamine. In addition to proneural genes (NGN2, MASH1), differentiated cells show expression of several genuine mesencephalic dopaminergic markers such as: LMX1A, LMX1B, GIRK2, ADH2, NURR1, PITX3, VMAT2 and DAT indicating that they retain their regional identity. Our data indicate that this cell line and its clonal derivatives may Constitute good candidates for the study of development and physiology of human dopaminergic neurons in vitro, and to develop tools for Parkinson's disease cell replacement preclinical research and drug testing. (C) 2009 Elsevier Inc. All rights reserved.
引用
收藏
页码:1860 / 1874
页数:15
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