, c6 x1 _" w" A, m( ` 5 |4 ^9 U2 x5 D9 r7 K7 [8 n% C # i+ E5 ~, W9 `8 Z- K 【摘要】 目的 研究应用超顺磁氧化铁（SPIO）标记神经干细胞(NSCs)移植治疗帕金森病（PD）大鼠脑部MRI的改变。方法 从大鼠胚胎脑中分离培养中脑NSCs，用脂质体转染法将SPIO标记NSCs；同时制作大鼠PD模型，SPIO标记的NSCs移植到PD大鼠右侧纹状体区，分别在移植后1周、2周、4周、6周、8周和10周给大鼠行MRI检查，2周、4周、6周、8周、10周时进行旋转行为评分，并与NSCs组和对照组比较。结果 脑内移植SPIO标记NSCs的PD大鼠1周后MRI检查显示在移植区呈低信号改变；移植后10周，在T2和T2*梯度回波仍可观察到移植区的低信号，同时移植区有混杂信号。NSCs移植大鼠的旋转行为评分在移植后2～10周明显少于对照组(均P " N6 u2 g) K9 c 【关键词】神经干细胞；超顺磁氧化铁；MRI；帕金森病；移植% v0 ^) f' }0 j
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Study the changes of brain MRI following transplantation of SPIO labeled neuralstem cells in treatment of Parkinsons disease in rat # h: |! w2 u2 {' `% M, L$ N' F2 I9 O8 @0 j8 m" P
YANG Lin, XIA Ying, ZHAO Hongyang, et al.4 S( D0 n0 x/ M) c
2 t& n) q. g. v: ] Department of Neurosurgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China# y; T3 X7 t" U- P
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$ ^0 p; o; ^! @: d7 |5 W# J Abstract：ObjectiveTo investigate the changes of brain MRI following transplantation of superparamagnetic iron oxide (SPIO) labeled neural stem cells (NSCs) in treatment of Parkinsons disease in rats.MethodsMutipotential NSCs from the midbrain of SD rats at embryonic day 14～16 were prepared. Labeled NSCs were achieved with the magnetic resonance contrast agent SPIO.Lipofectamine was used as transfection agent to achieve high transfer efficiencies. NSCs and SPIO NSCs were transplanted to the right striatal area of 6OHDA induced PD rat models. MRI scanning was performed at 1, 2, 4, 6, 8 and 10 weeks following transplantation and rotational behavior in response to amphetamine was tested at the same time.ResultsHypointense changes of MRI examination could be observed in the transplanted area of NSCs labeled by SPIO 1 week after transplantation in PD rats.10 weeks after transplantation, the hypointense signals and mixed signals were also found on T2 weighted and T2* gradient echo imaging. Rotational behavior in response to amphetamine showed a significant reduction in PD rats transplanted with NSCs labeled with SPIO compared with control group (all P # L# s2 \" E. u& e; _+ G. Y& V# J8 J* H8 [
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Key words：neural stem cells；superparamagnetic iron oxide；MRI；Parkinsons disease；transplantation4 M2 h. E! l1 [6 v: v
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多潜能的神经干细胞（NSCs）已经成为修复中枢神经系统损伤的细胞替代物，为神经系统疾病的治疗提供了一种新的途径［1］。但帕金森病(PD) NSCs移植治疗的临床应用还存在许多问题，特别是移植后如何区分移植的NSCs和宿主细胞［2］，目前仍缺乏有效的无创伤监测手段在体观察NSCs移植后的状况。本实验采用超顺磁氧化铁(SPIO)标记的NSCs对PD大鼠进行脑内移植治疗，并用MRI观察移植后NSCs的分布和迁徙情况。0 o; l* C; a$ l
) e9 r6 X0 m5 t* p% a6 w- v I 1材料与方法# N8 T/ u7 d* e. y; [5 c' X% I2 |3 P
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1.1材料孕14～16 d SD大鼠2只及成年SD大鼠（SPF级）20只，质量250～300 g，购自中国科学院上海分院实验动物中心。细胞培养试剂DMEM/F12、Neurobasal、人重组碱性成纤维细胞生长因子（hbFGF）、N2 supplement、B27 supplement购自Gibico BRL公司；鼠抗人神经巢蛋白（nestin）单克隆抗体(BD Pharmingen公司)；多聚赖氨酸（Sigma），脂质体Lipofectamine(Invitrogen公司)；Feridex（美国Advanced maganetic公司）。微量注射器(Hamilton)，SXP1手术显微镜(上海医用光学仪器厂)，荧光显微镜(LEICAL )，美国GE公司Signa Horizon LX 1.5 T超导磁共振成像系统，立体定向仪(Kopf)。 0 @+ c) _- I6 a, E- W! k ) b9 d7 |# c4 e! e4 E9 b5 [; l 1.2方法+ F) X. F# ^8 e o+ L9 R; p/ Y
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1.2.1中脑NSCs分离、培养和鉴定将怀孕的SD大鼠腹腔注射1％戊巴比妥钠（40 mg/kg）麻醉后，解剖显微镜下取出胚胎，在其中脑和间脑之间横断，以中脑曲为标志，从背侧用探针沿中线分开后，取腹侧约110 mm115 mm110 mm组织块，反复吹打成单细胞悬液，200目过滤，1500 r/min离心10 min，去除上清液，加入DMEM/F12培养液(含1%N2、2作者: 剑啸寒 时间: 2015-6-3 12:44