人脐血单个核细胞移植在缺氧缺血性脑损伤新生大鼠脑内的迁移及分化  

Migration and differentiation of human umbilical cord mononuclear cells in the brain of neonate rats following hypoxic-ischemic brain damage

作　　者：王晓莉[1] 赵岩松[2] 李耀武[1] 徐敏[1] 

机构地区：[1]潍坊医学院医学影像学系分子影像学研究中心,山东省潍坊市261042 [2]潍坊医学院眼科教研室,山东省潍坊市261042

出　　处：《中国组织工程研究与临床康复》2009年第32期6259-6262,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：潍坊医学院博士启动基金(KB200802);山东省自然科学基金项目(Y2007C084);山东省教育厅科技计划项目(J07YE02)~~

摘　　要：背景:研究发现脐血单个核细胞脑内移植后可以存活,并减轻缺氧缺血性脑损伤程度,显著改善脑损伤大鼠的远期行为学,但作用机制仍未阐明。目的:将人脐血单个核细胞经侧脑室移植入缺氧缺血性脑损伤新生大鼠脑内,观察植入细胞在脑内的迁移与分化。设计、时间及地点:细胞学体内观察,于2007-12/2008-08在潍坊医学院分子影像学研究中心完成。材料:脐血来源于健康、足月妊娠产妇,由潍坊医学院附属医院产科提供。清洁级健康7d龄SD新生大鼠50只,由山东省中医药大学动物中心提供。方法:Ficoll法体外分离培养人脐血单个核细胞,移植前3d行BrdU标记。50只大鼠均采用Rice-Vannucci法复制缺氧缺血性脑损伤模型,造模后24h,在左侧侧脑室(AP:-0.5mm,ML:-2mm,DV:-2mm)注入脐血单个核细胞悬液,2μL/点,共1×105个活细胞。分别于细胞移植后24h,3d,7d,14d,28d取材制备脑组织切片,10只/时间点。主要观察指标:采用免疫荧光双标法检测移植脐血单个核细胞的脑内迁移、神经干细胞的表达、分化情况。结果:50只大鼠均进入结果分析。①移植后24h侧脑室内可见BrdU+细胞;移植后3,7d移植细胞迁移到室管膜下区;移植后14,28d移植细胞迁移到大脑皮质及海马。②移植后24h侧脑室内可见BrdU+nestin+细胞;移植后3d室管膜下区出现BrdU+nestin+细胞;移植后7dBrdU+nestin+细胞数增加;移植后14dBrdU+nestin+细胞数下降。③移植后14d大脑皮质、海马开始出现BrdU+NSE+细胞与BrdU+GFAP+细胞,移植后28d双阳性细胞数进一步增加,BrdU+NSE+细胞及BrdU+GFAP+细胞占BrdU+的比例也增加。结论:脐血单个核细胞侧脑室移植入脑内后可以继续存活,并迁移到大脑皮质与海马部位,分化为成熟的神经元与神经胶质细胞。BACKGROUND： Umbilical cord blood mononuclear cell （UMNC） transplantation in the brain can survive, reduce brain damage induced by hypoxia and ischemia, significantly improve long-term ethology in rats following brain damage, but the mechanisms are still unclear. OBJECTIVE： To investigate the migration and differentiation of human umbilical cord mononuclear cells after their transplantation to the brain via lateral ventricle in rats with brain damage induced by hypoxia and ischemia. DESIGN, TIME AND SETTING： The in vivo cytological study was performed at the Molecular Imaging Center, Weifang Medical College from December 2007 to August 2008. MATERIALS： Umbilical cord blood was obtained from healthy full-term pregnant women at the Department of Obstetrics, Hospital Affiliated to Weifang Medical College. A total of 50 clean healthy Sprague Dawley neonatal rats aged 7 days were supplied by Animal Center, Shandong University of Traditional Chinese Medicine. METHODS： UMNCs were obtained from human cord blood in vitro by Ficoll method, and labeled by BrdU 3 days prior to transplantation. A total of 50 rat models of hypoxic-ischemic brain damage were established by Rice-Vannucci method. UMNC suspension was infused into the left lateral ventricle （AP： -0.5 mm, ML： 2 mm, DV： -2 mm） 24 hours following model establishment, 2μL/point, totally 1×10^5 living cells. Brain tissue sections were prepared 24 hours, 3, 7, 14 and 28 days following transplantation, 10 rats per time point. MAIN OUTCOME MEASURES： Migration of UMNCs, expression and differentiation of neural stem cells in the brain were measured using double immunofluorescent staining. RESULTS： All the 50 rats were involved in final analysis.①24 hours after transplantation, BrdU^＋ cells were seen in the lateral ventricular; 3 and 7 days after transplantation, the BrdU^＋ cells were observed in the subventricular zone; 14 and 28 days after transplantation, the BrdU^＋ cells migrated to the cortex and hippocampus.②24 hours after transpla

关 键 词：脐血单个核细胞 移植 缺氧缺血 分化 

分 类 号：R394.2[医药卫生—医学遗传学]