永生化神经前体细胞系hNPC-TERT体内外D_2受体的表达  

D_2 receptor expression on immortalized human neural progenitor cell line hNPC-TERT in vitro and in vivo

作　　者：丁为民[1] 田嘉禾[2] 杨晓华[3] 白金柱[4] 沈丽[4] 张锦明[2] 

机构地区：[1]南方医科大学珠江医院肿瘤中心,广东广州510282 [2]中国人民解放军总医院核医学科,北京100853 [3]南方医科大学珠江医院神经内科,广东广州510282 [4]北京大学干细胞研究中心,北京100083

出　　处：《南方医科大学学报》2007年第6期795-798,共4页Journal of Southern Medical University

基　　金：国家863高技术研究发展计划资助项目(2002AA205081)~~

摘　　要：目的验证神经前体细胞系hNPC-TERT移植入动物中枢神经系统前后多巴胺D2受体的表达。方法免疫荧光定性鉴定体外培养hNPC-TERT的D2受体表达,并进一步应用放射受体分析方法测定hNPC-TERT细胞表达D2受体数量。实验组兔脊髓内移植3×106hNPC-TERT,对照组动物脊髓内移植同等数量HeLa细胞,移植后第7天,以11C-raclopride为示踪剂,移植动物活体或处死动物剥离出脊髓进行D2受体PET显像,并行移植段脊髓切片免疫荧光染色,验证hNPC-TERT在体内的D2受体表达。结果免疫荧光和放射受体分析显示体外培养hNPC-TERT细胞表面表达大量D2受体(Bmax=8×104)。移植入兔脊髓后,D2受体显像显示移植部位有明显的放射性浓聚,经ROI半定量分析,与对照组差异显著(P=0.0017),离体脊髓PET显像、组织免疫荧光进一步证实了活体显像结果。结论神经前体细胞系hNPC-TERT高表达多巴胺D2受体,并且在移植入体内后仍保持该特性,具有移植治疗帕金森综合征等D2受体缺乏性疾病的潜力。Objective To observe D2 receptor expression on human neural progenitor cell line hNPC-TERT before and after transplantation into rabbit central nervous system. Methods D2 receptor expression on cultured hNPC-TERT cells was verified and quantitatively analyzed with immunofluorescence assay and receptor radio ligand binding assay, respectively. 3×10^6 hNPC-TERT cells were implanted in the spinal cord of New Zealand rabbit with HeLa cells as the control. Two days after implantation, positron-emission tomography （PET） scan with ^11C-raclopride as the radiotracer was performed in the living animals or for the isolated spinal cords, and cryosections of the spinal cord containing the implanted cells were prepared for imrnunofluorescence assay. Results Cultured hNPC-TERT cells showed high expression of D2 receptor （Bmax=8× 10^4）. PET scans of the rabbits identified visible radioactive accumulations at the site where hNPC-TERT ceils were implanted but not at the site of HeLa cell implantation. Region of interest analysis showed a significant difference between the two cells in the maximal standard uptake value at the cell implantation sites. The results were further confirmed with ex vivo PET imaging of the spinal cord and tissue immunofluorescence assay. Conclusion Human neural progenitor cells hNPC-TERT highly express dopamine D2 receptors and retain this capacity after implantation into the spinal cord, suggesting their potential for treatment of such nerve system disease as Parkinson syndrome.

关 键 词：神经前体细胞 hNPC-TERT 移植 D2受体 

分 类 号：R651[医药卫生—外科学]