胆碱能前体细胞移植入β-淀粉样前体蛋白转基因小鼠脑内的存活和迁移  

Survival and Migration of Cholinergic Precursor Cells Transplanted into APP Transgenic Mice

作　　者：曹敬丽[1] 王雅杰[1] 

机构地区：[1]首都医科大学附属北京天坛医院临床医学研究实验室,北京100050

出　　处：《标记免疫分析与临床》2015年第9期912-915,共4页Labeled Immunoassays and Clinical Medicine

基　　金：国家自然科学基金(81572474);北京市自然科学基金(7144209)

摘　　要：目的观察新生小鼠海马原代培养的胆碱能神经前体细胞移植入淀粉样前体蛋白(amyloid precusor protein,APP)转基因小鼠脑内以后的存活和迁移状况,以探索胆碱能神经前体细胞对阿尔茨海默病的治疗前景。方法取新生24h以内增强型绿色荧光蛋白(enhanced green fluorescent protein,EGFP)转基因小鼠,以原代细胞分离培养方法获得海马胆碱能神经前体细胞,以前体细胞特异性标记抗巢蛋白(Nestin)和胆碱能细胞特异性标记胆碱乙酰基转移酶(choline acetyltransferase,Ch AT)做细胞免疫组化鉴定细胞特性。细胞培养成功后,收集细胞,在小鼠脑立体定位仪的固定下,参照小鼠脑立体定位图谱对APP转基因小鼠施行脑内注射。移植1周以后灌注取移植鼠脑组织行冰冻切片,在共聚焦显微镜下通过外源细胞的绿色自发荧光观察移植细胞的存活和迁移情况。结果在移植位点,外源性绿色荧光细胞存活良好并以移植点为中心向外迁移,且呈继续迁移的趋势。结论新生小鼠海马原代细胞培养以后呈现神经前体细胞的特性并且胆碱能阳性,移植入APP转基因小鼠脑内以后,这些胆碱能阳性的神经前体细胞存活良好并呈现较强迁移能力,为进一步探索该细胞对阿尔茨海默病的治疗可能性提供了依据。Objective To explore the survival and migration of cholinergic precursor cells transplanted into APP transgenic mice and to evaluate the possibility of alzheimer＇ s disease therapy with cholinergic precursor cells. Methods Newborn EGFP transgenic mice were adopted for primary cholinergic precursor cells culture, the Nestin and ChAT were used for immunohistochemistry characterization. Stereotaxic Coordinates and anterior- posterior were applicated for microinjection. One week after transplantation, the transplanted mice were perfused and the brains were removed for frozen section, then confocal microscope was adopted to observe the survival and migration of the transplanted cells and the results were collected. Results In the cell culture, the primary cells showed both Nestin and ChAT positive. One week after transplantation, the transplanted EGFP positive cells at the injection site survived well and showed strong ability of migration and tended to migrate continuously. Conclusion In the cell culture, the primary hippocampus cells showed characterization of neural precursor and cholinergic cells. After transplanted into the brain of APP transgenic mice, the cells survived well and showed strong ability for migration. This implicated that cholinergic precursor cells maybe an effective therapy for Alzheimer＇ s Disease.

关 键 词：胆碱能前体细胞 海马 细胞移植 绿色荧光蛋白 淀粉样前体蛋白 

分 类 号：R749.16[医药卫生—神经病学与精神病学]