中国组织工程研究与临床康复
中國組織工程研究與臨床康複
중국조직공정연구여림상강복
JOURNAL OF CLINICAL REHABILITATIVE TISSUE ENGINEERING RESEARCH
2008年
47期
9394-9400
,共7页
汤锋武%修波%范存刚%崔志强%萧凯%韩中朝
湯鋒武%脩波%範存剛%崔誌彊%蕭凱%韓中朝
탕봉무%수파%범존강%최지강%소개%한중조
CD133%痴呆%细胞移植%认知功能%存活
CD133%癡呆%細胞移植%認知功能%存活
CD133%치태%세포이식%인지공능%존활
背景:脐血中富含早期干细胞的一个亚群CD133+细胞,该亚群是具有广泛的增生和分化潜能的原始细胞,被认为具有向神经细胞分化的潜能.目的:假设脐血CD133+细胞对改善痴呆小鼠的认知功能和存活有使用价值,检测脐血CD133+细胞移植后痴呆鼠的认知和存活功能的变化,拟予验证.设计、时间和单位:完全随机区组设计的动物实验,于2005-09/2007-01在天津血液学研究所完成.材料:选用48只雄性APP695转基因小鼠,随机分为3组:对照组(n=8)、CD133+细胞移植组(n=20)和CD133细胞移植组(n=20).方法:对照组小鼠经脑室注射10μ L磷酸缓冲液(PBS),CD133+细胞移植组和CD133细胞移植组分别经脑室注射10 μ LCD133+(5×104/μ L)和CD133(5×104/μL)脐血细胞移植.主要观察指标:以水迷宫实验评价转基因小鼠在细胞移植后的认知功能,并记录移植后小鼠的存活时间.以Dil荧光标记法和免疫组化检测移植细胞的分化类型.结果:在移植后30 d,CD133+细胞移植组小鼠的认知功能明显好于CD133细胞移植组和对照组,差异有显著性意义(P＜0.05),这种改善效果在移植后180 d仍然存在(P＜0.05).CD133+细胞移植组小鼠平均存活时间比CD133细胞移植组和对照组明显延长,差异有显著性意义(P＜0.05).标记有Dil的脐血细胞在移植后30 d已经迁移到多个脑区,其中主要在双侧顶叶皮质和海马区,并且能够表达神经细胞标志Ⅲ型β微管蛋白、神经微丝、神经烯醇化酶和胶质酸性蛋白.CD133细胞移植组脑切片内,Dil标记的脐血源细胞主要分布在侧脑室及其周围,很少能检测到阳性胶质酸性蛋白,Ⅲ型β微管蛋白,或神经烯醇化酶的脐血细胞.移植后30 d,CD133+细胞移植组Dil标记的脐血细胞表达Ⅲ型β微管蛋白的百分率明显高于180 d,表达神经烯醇化酶的脐血细胞百分率明显低于180 d,差异均有显著性意义(P＜0.01).结论:实验结果提示,移植CD133+细胞后出现的对认知功能改善和对存活时间延长的作用,可能主要归功于移植细胞对功能不良细胞的替代或者对神经环路的改善.
揹景:臍血中富含早期榦細胞的一箇亞群CD133+細胞,該亞群是具有廣汎的增生和分化潛能的原始細胞,被認為具有嚮神經細胞分化的潛能.目的:假設臍血CD133+細胞對改善癡呆小鼠的認知功能和存活有使用價值,檢測臍血CD133+細胞移植後癡呆鼠的認知和存活功能的變化,擬予驗證.設計、時間和單位:完全隨機區組設計的動物實驗,于2005-09/2007-01在天津血液學研究所完成.材料:選用48隻雄性APP695轉基因小鼠,隨機分為3組:對照組(n=8)、CD133+細胞移植組(n=20)和CD133細胞移植組(n=20).方法:對照組小鼠經腦室註射10μ L燐痠緩遲液(PBS),CD133+細胞移植組和CD133細胞移植組分彆經腦室註射10 μ LCD133+(5×104/μ L)和CD133(5×104/μL)臍血細胞移植.主要觀察指標:以水迷宮實驗評價轉基因小鼠在細胞移植後的認知功能,併記錄移植後小鼠的存活時間.以Dil熒光標記法和免疫組化檢測移植細胞的分化類型.結果:在移植後30 d,CD133+細胞移植組小鼠的認知功能明顯好于CD133細胞移植組和對照組,差異有顯著性意義(P＜0.05),這種改善效果在移植後180 d仍然存在(P＜0.05).CD133+細胞移植組小鼠平均存活時間比CD133細胞移植組和對照組明顯延長,差異有顯著性意義(P＜0.05).標記有Dil的臍血細胞在移植後30 d已經遷移到多箇腦區,其中主要在雙側頂葉皮質和海馬區,併且能夠錶達神經細胞標誌Ⅲ型β微管蛋白、神經微絲、神經烯醇化酶和膠質痠性蛋白.CD133細胞移植組腦切片內,Dil標記的臍血源細胞主要分佈在側腦室及其週圍,很少能檢測到暘性膠質痠性蛋白,Ⅲ型β微管蛋白,或神經烯醇化酶的臍血細胞.移植後30 d,CD133+細胞移植組Dil標記的臍血細胞錶達Ⅲ型β微管蛋白的百分率明顯高于180 d,錶達神經烯醇化酶的臍血細胞百分率明顯低于180 d,差異均有顯著性意義(P＜0.01).結論:實驗結果提示,移植CD133+細胞後齣現的對認知功能改善和對存活時間延長的作用,可能主要歸功于移植細胞對功能不良細胞的替代或者對神經環路的改善.
배경:제혈중부함조기간세포적일개아군CD133+세포,해아군시구유엄범적증생화분화잠능적원시세포,피인위구유향신경세포분화적잠능.목적:가설제혈CD133+세포대개선치태소서적인지공능화존활유사용개치,검측제혈CD133+세포이식후치태서적인지화존활공능적변화,의여험증.설계、시간화단위:완전수궤구조설계적동물실험,우2005-09/2007-01재천진혈액학연구소완성.재료:선용48지웅성APP695전기인소서,수궤분위3조:대조조(n=8)、CD133+세포이식조(n=20)화CD133세포이식조(n=20).방법:대조조소서경뇌실주사10μ L린산완충액(PBS),CD133+세포이식조화CD133세포이식조분별경뇌실주사10 μ LCD133+(5×104/μ L)화CD133(5×104/μL)제혈세포이식.주요관찰지표:이수미궁실험평개전기인소서재세포이식후적인지공능,병기록이식후소서적존활시간.이Dil형광표기법화면역조화검측이식세포적분화류형.결과:재이식후30 d,CD133+세포이식조소서적인지공능명현호우CD133세포이식조화대조조,차이유현저성의의(P＜0.05),저충개선효과재이식후180 d잉연존재(P＜0.05).CD133+세포이식조소서평균존활시간비CD133세포이식조화대조조명현연장,차이유현저성의의(P＜0.05).표기유Dil적제혈세포재이식후30 d이경천이도다개뇌구,기중주요재쌍측정협피질화해마구,병차능구표체신경세포표지Ⅲ형β미관단백、신경미사、신경희순화매화효질산성단백.CD133세포이식조뇌절편내,Dil표기적제혈원세포주요분포재측뇌실급기주위,흔소능검측도양성효질산성단백,Ⅲ형β미관단백,혹신경희순화매적제혈세포.이식후30 d,CD133+세포이식조Dil표기적제혈세포표체Ⅲ형β미관단백적백분솔명현고우180 d,표체신경희순화매적제혈세포백분솔명현저우180 d,차이균유현저성의의(P＜0.01).결론:실험결과제시,이식CD133+세포후출현적대인지공능개선화대존활시간연장적작용,가능주요귀공우이식세포대공능불량세포적체대혹자대신경배로적개선.
BACKGROUND:Human umbilical cord blood (CB)-derived CD133+ cells are a minority population of primitive cells with extensive proliferation and differentiation potentials,which are considered to have ability of neural differentiation.OBJECTIVE:We hypothesized a possible application of CB CD133+ cells in the cognitive and survival function of mice with dementia,the present study observed the changes of the cognitive function and survival of amyloid precursor protein(APP)transgenic mice after CB CD 133+ cells transplantation to verify the above assumption.DESIGN,TIME AND SETTING:A completely randomized block design of animal experiments was performed in the Hematology Institute of Tianjin Hematology Hospital from September 2005 to December 2007.MATERIALS:Forty-eight eight-month-old male APP 695 transgenic C57BL/6 (BDF1/KM) mice were selected in this experiments All mice were divided randomly into three groups:control group (n=8),CD133+ transplantation group (n=20) and CD133 transplantation group (n=20).METHODS:Mice in control groups received an intraventricular injection of 10 μL phosphate buffered saline (PBS).The transgenic mice that received an intraventricular injection of 10 μL CD133+ (5×104/μL) and CD133 CB cells (5×104/μL) respectively.MAIN OUTCOME MEASURES:Radial ann water maze (RAWM) was used to evaluate cognitive function of the mice and the survival days of mice in different groups were recorded,lmmunohistochemical assessments and Dil Fluorescence labeled way was used to detect the differentiation phenotype of transplanted cells.RESULTS:The cognitive function of the mice in CD133+ transplantation group was significantly improved compared with the mice in CD 133- transplantation and control groups both 30 and 180 days after transplantation (P＜0.05).The mean survival time of the mice in CD133+ transplantation group was significantly increased compared with CD133 transplantin group and control group (P＜0.05).It was observed that the transplantation CB CD133+ cells labeled with Dil migrated into several brain regions at day 30 post-transplantation.These cells were stained for human βⅢ-tubulin,neuralfilement(NF),neuron specific enolase (NSE),and glial fibriliary acidic protein(GFAP).However,in the brain of mice that received CD133 cells transplantation,CB cells were distributed mainly in and around the lateral ventricle at day 30 and 180 post-transplantation and GFAP-,βⅢ-tubulin- and NSE-positive cells were rarely detected.After intraventricular transplantation of CB CD133+ cells,the percentage of transplanted Dil-labeled CB cells expressing βⅢ-tubulin was significant higher at day 30 than at day 180,and the percentage of CB cells expressing NSE was significant lower at day 30 than that at day 180 (both P＜0.01).The percentage of CB cells expressing GFAP was relatively constant between the days 30 and 180 after transplantation (P＞0.05).CONCLUSION:The result of this experiment suggested that the cognitive and survival function improvement achieved by transplantation of CB CD133+ cells is mainly due to a replacement of dysfunctional cells or augmentation of neural circuit by CB CD133+ cells transplantation.
