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Cell Journal، جلد ۱۵، شماره Suppl ۱، صفحات ۴۵-۴۵
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عنوان انگلیسی Ps-58: Isolation of A Stably Transformed PC12 Cell Line Expressing Human ANXA1 cDNA
چکیده انگلیسی مقاله Objective: Parkinson's disease (PD) is known as a degenerative disorder of the central nervous system. In this disease, the death of dopaminergic neurons in the substantia nigra, a region of the midbrain is occurred. Annexin A1 (AnxA1) is reported as an anti-inflammatory factor. ANXA1 might play a neuroprotective role in PD by acting as an anti-inflammatory mediator. The aim of this study was isolation of PC12 cell line overexpressing ANXA1. Materials and Methods: At first step, cDNA of ANXA1was prepared from neutrophils and then inserted to PGL268 vector to produce recombinant PGL/ANXA1 vector. The accuracy of cloning was confirmed by further sequencing. Transfection was carried out in the dopaminergic neuronal cell line, PC12 (a cell line derived from a pheochromocytoma of the rat adrenal medulla), using of lipofectamin LTX and after 48 hours EGFP reporter was detected. Furthermore, PC12 cells expressing ANXA1 gene were selected followed by G418 treatment after 2 weeks. Expression of ANXA1 was evaluated by real time PCR, western blot and flow cytometry techniques. Results: The results indicated strong expression of AnXA1 in PC12 cells. Conclusion: PC12 cells containing ANXA1 DNA could be further used as an in vitro model for evaluation of ANXA1 Anti-inflammatory effects.
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نشانی اینترنتی http://celljournal.org/journal/article/abstract/862
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