Document Detail


Establishment of an immortalized GABAergic neuronal progenitor cell line from embryonic ventral mesencephalon in the rat.
MedLine Citation:
PMID:  18407253     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Effective cell replacement therapies for neurological disease require neuron-restricted precursors as grafted cells. The problem of obtaining sufficient grafts for transplantation can be resolved by creating an appropriate immortalized cell line. In the present study, a thermally controlled immortalized GABAergic neuronal progenitor cell line (RMNE6) was established from E13 rat ventral mesencephalon cells immortalized using the temperature-sensitive mutant of SV40 large T antigen (ts-TAg). RMNE6 cells proliferated rapidly and expressed a neuron-like phenotype at the permissive temperature (33 degrees C), but eventually stopped growing at the non-permissive temperature (39 degrees C). Expression of the neuronal markers PSA-NCAM, beta-tubulin III and MAP2 by RMNE6 cells was confirmed by RT-PCR or immunocytochemistry. Furthermore, these cells exhibited functional GABAergic neuron properties, as evidenced by the expression of glutamate decarboxylase (GAD) as well as the synthesis and release of the neurotransmitter GABA in a calcium-dependent manner. Moreover, RMNE6 cells spontaneously expressed and secreted several neurotrophic factors, such as NGF, BDNF, NT-3, NT-4/5, and GDNF. The cells survived well and kept expression of SV40 Tag, GAD65/67 and GABA in the striatum, at least 28 days after being transplanted in the rat brain. Tumorigenesis assays confirmed the safety of the immortalized cell line in vivo. Taken together, the results support the use of RMNE6 cells as an ideal cell model for transplantation research aimed at the treatment and prevention of neurodegenerative disease.
Authors:
Beibei Wang; Xifeng Zou; Haiyan Zhang; Deyi Duan; Lili Ju; Xiaohua Jiang; Xiaohong Sun; Chunli Zhao; Huanying Zhao; Jin Guo; Changlei Xu; Erjing Gao; Qunyuan Xu
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2008-03-04
Journal Detail:
Title:  Brain research     Volume:  1210     ISSN:  0006-8993     ISO Abbreviation:  Brain Res.     Publication Date:  2008 May 
Date Detail:
Created Date:  2008-05-12     Completed Date:  2008-08-26     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0045503     Medline TA:  Brain Res     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  63-75     Citation Subset:  IM    
Affiliation:
Beijing Institute for Neuroscience, Capital Medical University, Beijing Center of Neural Regeneration and Repair, Key Laboratory of Neurodegenerative diseases of Ministry of Education of China, Beijing 100069, China.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antigens, Polyomavirus Transforming / genetics
Biological Markers / metabolism
Brain Tissue Transplantation / methods*
Cell Culture Techniques / methods
Cell Differentiation / physiology
Cell Line, Transformed*
Cell Proliferation
Cells, Cultured
Female
Glutamate Decarboxylase / metabolism
Male
Mesencephalon / cytology*,  embryology,  metabolism
Mice
Mice, Inbred BALB C
Nerve Growth Factors / metabolism
Nerve Tissue Proteins / metabolism
Neurons / cytology*,  metabolism
Rats
Rats, Sprague-Dawley
Stem Cells / cytology*,  metabolism
Temperature
Transplantation, Heterologous
Transplantation, Homologous
gamma-Aminobutyric Acid / metabolism*
Chemical
Reg. No./Substance:
0/Antigens, Polyomavirus Transforming; 0/Biological Markers; 0/Nerve Growth Factors; 0/Nerve Tissue Proteins; 56-12-2/gamma-Aminobutyric Acid; EC 4.1.1.15/Glutamate Decarboxylase

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