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1.
脑出血大鼠脑内神经干细胞移植的研究   总被引:4,自引:0,他引:4  
目的分离并克隆新生大鼠神经干细胞,研究其移植入脑出血大鼠脑内的生物学特征,了解神经干细胞移植治疗脑出血的可行性.方法用尾状核注射Ⅶ型胶原酶制作脑出血模型,从Wistar新生大鼠脑室下区分离并克隆神经干细胞,经Brdu(5-溴脱氧尿嘧啶)掺入标记后移植入脑出血同侧的侧脑室或脑出血对侧的尾状核中.经免疫组织化学鉴定了解移植细胞在大鼠脑内的生存、迁移及分化情况.结果将稳定培养的神经干细胞移植入脑出血大鼠脑内,发现移植后4 d移植细胞仍存在.侧脑室移植组中移植细胞多在侧脑室周边区存在,尾状核移植组可见移植细胞开始向对侧迁移.免疫荧光双标证实细胞大多分化成神经元,少部分分化成胶质细胞.结论神经干细胞移植入脑出血大鼠脑内后能够存活,并能有效地穿过室管膜和向脑出血部位迁移.移植细胞在脑内大部分分化成神经元,少部分分化成胶质细胞.  相似文献   

2.
背景:目前认为直接进行神经干细胞移植后细胞虽能存活,但是只分化成神经胶质细胞,并不能分化成有功能的神经元。 目的:探讨维甲酸诱导对胚胎大鼠脑海马神经干细胞向神经元分化的作用。 设计、时间及地点:细胞学体外实验,于2008-09/2009-02在辽宁医学院科技实验楼完成。 材料:胚龄13.5 d的SD大鼠由辽宁医学院实验动物中心提供。 方法:分离胎鼠脑海马组织,胰蛋白酶消化法体外培养获得神经干细胞。将原代和传代细胞以1×107 L-1接种到培养孔中,分别进行常规贴壁分化培养和维甲酸诱导分化培养。 主要观察指标:神经干细胞的鉴定,光镜及免疫组化检测神经干细胞诱导分化结果。 结果:免疫组织化学检测结果显示,原代和传代后得到的神经干细胞团均呈巢蛋白阳性。常规贴壁分化培养7 d后,神经元多呈椭圆型和近似三角形,胞体大,细胞边缘清楚,胞体上有多个突起;而维甲酸诱导分化培养后,神经元数量增加,形态清楚,但细胞胞体上突起较少。与常规贴壁分化培养比较,维甲酸诱导分化培养后神经干细胞向神经元分化率明显升高(P < 0.01),神经干细胞向神经胶质细胞分化率明显降低(P < 0.01)。 结论:从大鼠胚胎脑海马组织中分离得到可自我复制和多向分化的神经干细胞,维甲酸体外诱导后可以增加其向神经元方向分化的比例。  相似文献   

3.
目的 观察转染绿色荧光蛋白(GFP)的大鼠脊髓神经干细胞移植于半横断脊髓损伤处的体内外分化情况.方法 将表达GFP的慢病毒载体转染胎鼠脊髓神经干细胞,体外用10%胎牛血清诱导分化.转染后的神经干细胞与PLGA支架移植于大鼠半横断脊髓损伤处,术后1个月和3个月取材,行GFAP、NF和CNP免疫荧光染色.结果 转染GFP的神经干细胞球表达强烈的绿色荧光,体外分化可见GFAP/GFP、NF/GFP和CNP/GFP双阳性细胞,GFAP/GFP双阳性细胞明显多于其他两种.移植后3个月,GFP阳性细胞在脊髓内明显减少,可见少数GFAP/GFP和CNP/GFP舣阳性细胞,未见NF/GFP双阳性细胞.结论 转染GFP的神经干细胞可在体外增殖和分化,但大部分分化成胶质细胞.移植于急性期脊髓损伤处的神经干细胞不被诱导分化成神经元样细胞,可被诱导分化成神经胶质细胞.  相似文献   

4.
目的对比神经干细胞(NSCs)和嗅鞘细胞(OECs)联合移植与NSCs移植治疗大鼠脑出血的疗效。方法制作大鼠脑出血模型,记录细胞移植后对照组和移植组(NSCs组、NSCsOECs组)大鼠运动功能,采用免疫组化观察移植细胞在体内存活、分化和迁徙的情况。结果NSCsOECs组和NSCs组移植NSCs分别有14.19%和2.96%分化为神经元,差异有统计学意义(χ2=154.79,P<0.01),30d时神经功能缺损评分NSCs组为1.60±0.13,NSCsOECs组为1.01±0.11,差异有统计学意义(P<0.05),OECs移植后能存活、迁徙,NSCs移植后能存活、分化为神经元、星形和少突胶质细胞,并向损伤区迁徙。结论OECsNSCs移植改善脑出血大鼠运动功能的疗效优于单纯NSCs移植。  相似文献   

5.
背景:外源性神经干细胞具有神经修复作用,可能对脑出血后的神经功能恢复起到一定的作用。 目的:观察胎鼠神经干细胞的体外生长、分化及移植到脑出血大鼠后的存活、迁徙、分化情况,探讨神经干细胞对脑出血模型大鼠受损神经功能的修复作用。 设计:完全随机分组设计,对照动物实验。 单位:复旦大学附属华山医院神经外科 材料:选用健康雄性成年SD大鼠18只为受体,体质量280~320 g,由中国科学院上海实验动物中心提供。实验用鼠抗BrdU为Neomarkers产品, 鼠抗胶质纤维酸性蛋白和兔抗微管相关蛋白2 为Chemicon产品。 方法:实验于2006-02/12在复旦大学附属上海医学院解剖组胚实验室完成。从胎龄14 d的胎鼠海马中分离、培养、鉴定神经干细胞。16只受体SD大鼠被随机分为3组:对照组,PBS组和移植神经干细胞组。均通过尾状核内注射自体动脉血制作大鼠脑出血模型。移植NSC组在造模后30 min在血肿腔周围四点分别移植浓度为2×1011 L-1神经干细胞悬液5μL;PBS组于相同时间点在脑内相同部位注射PBS;PBS和神经干细胞的移植方法同自体血的移植方法。对照组大鼠在造模后30 min只造成四点损伤,不注射任何物质。 主要观察指标:在造模后立即,1,3,5,14,21,28 d采用前肢评分和转身评分对大鼠神经功能进行评估。大鼠于造模后28 d麻醉后取脑,并通过双标胶质纤维酸性蛋白、微管相关蛋白2、BrdU免疫组化来检测移植入脑的神经干细胞在体内的分化情况。 结果:①神经功能评分:造模后5 d,各组差异无显著性意义(P > 0.05)。造模后14~28 d,干细胞移植组较其他3组明显改善(P < 0.05)。②脑组织切片双免疫组织学双标染色结果:干细胞移植组血肿周围凋亡细胞少于PBS组。受体大鼠脑组织切片显示有BrdU, 微管相关蛋白2,胶质纤维酸性蛋白阳性细胞,说明神经干细胞可以在宿主脑内存活、迁徙和分化,可以分化为神经元样细胞和神经胶质样细胞。 结论:神经干细胞移植可能通过分化为神经元样细胞和神经胶质细胞促进大鼠脑出血的神经功能恢复。  相似文献   

6.
目的 研究胚胎神经干细胞出血性脑卒中大鼠脑内移植对神经功能缺损改善作用.方法 30只SD大鼠随机分配到单纯脑出血组(A,10只)、培养液移植组(B,10只)及NSCs移植组(C,10只).制作脑出血模型后3d,C组将神经于细胞经Hoechst标记后移植到病灶区,B组注射等量培养液.通过移植前后神经功能评分检测神经功能缺损修复情况,以及冰冻切片后组织免疫荧光检测移植神经干细胞的分化情况.结果 实验中分离、培养的神经干细胞经标记移植人大鼠脑内后,经免疫荧光检测证实能够在血肿腔周边分化成神经元和星形胶质细胞.移植术后2周内3组间大鼠神经功能改善无明显差异.但从移植术后第21天到第28天,神经干细胞移植组大鼠的神经功能改善显著好于培养液移植组和单纯脑出血组P<0.001,有统计学意义.结论 神经干细胞移植入脑出血大鼠脑内后能够存活并分化为神经元及星形胶质细胞,促进改善大鼠的神经功能,是一种很有发展前途的治疗方法,值得进一步深入研究.  相似文献   

7.
血清和雪旺氏细胞诱导大鼠胚胎神经干细胞分化的比较   总被引:1,自引:0,他引:1  
目的 比较血清和雪旺氏细胞诱导大鼠胚胎神经干细胞分化的差异。方法 分别采用血清和与雪旺氏细胞共培养的方法诱导大鼠胚胎神经干细胞分化,应用相差显微镜和免疫荧光染色的方法对其进行观察和比较。结果 两种方法都能够诱导绝大多数神经干细胞分化成神经元,少量分化成星形胶质细胞和少突胶质细胞。虽然后一种方法诱导干细胞分化的进程比前一种方法要慢,但细胞形态学上更接近发育成熟的神经元。结论 雪旺氏细胞的分泌物不仅能够诱导共培养的神经干细胞分化,而且使其分化更加成熟。  相似文献   

8.
体内外不同环境对大鼠胚胎神经干细胞分化的影响   总被引:6,自引:2,他引:4  
目的 探讨体内外不同环境条件对大鼠胚胎神经干细胞分化影响。方法 孕龄16天的大鼠胚胎神经干细胞体外扩增后移植至大鼠脑内尾状核出血部位;同时体外将其与新生大鼠Schwann细胞共培养,应用免疫组织化学和免疫荧光方法分另检测神经细胞、星形胶质细胞及少突胶质细胞特异性标志蛋白tubulin-β、MAP2,GFAP和GalC的表达。结果 移植到脑内的BrdU阳性细胞环绕脑出血区域分布,大部分细胞GFAP阳性,少部分细胞tubulin-β或GalC阳性;而与Schwann细胞共培养后,神经干细胞大部分呈tubulin-β和MAP2免疫荧光阳性,少部分呈GFAP或GalC免疫荧光阳性。结论 脑内尾状核出血环境诱导神经干细胞主要分化成神经胶质细胞;而体外与Schwann细胞共培养主要诱导其分化成神经细胞。  相似文献   

9.
目的:观察胎鼠神经干细胞体外的生长、分化及移植到脑出血大鼠后的存活、迁徙、分化情况。研究神经干细胞对脑出血后神经功能损害的可能修复作用。方法:通过尾状核内注射自体动脉血制作大鼠脑出血模型。分离、培养大鼠胎鼠神经干细胞并移植于成年大鼠尾状核,对大鼠脑出血后的神经恢复情况进行功能评价。结果:神经干细胞可以在宿主脑内存活、迁徙和分化,神经干细胞移植组大鼠运动功能较对照组明显改善。结论:神经干细胞移植能促进改善大鼠脑出血的神经功能恢复。  相似文献   

10.
大鼠胚胎神经干细胞的分离、培养和鉴定   总被引:1,自引:0,他引:1  
目的 探讨大鼠胚胎神经干细胞体外分离培养增殖及分化,为进一步的实验研究提供基础。方法 原代培养.培养细胞生长状况观察用无血清培养技术进行培养、传代和鉴定。诱导分化后采用SABC法对分化的细胞进行神经元特异性烯醇化酶(NSE)、神经胶质酸性蛋白(GFAP)检测以作细胞鉴定。结果 成功培养出大鼠胚胎神经干细胞,了解其生长规律,并对其进行传代、冻存及复苏,培养的细胞能分化为神经元细胞和神经胶质细胞。结论 大鼠胚胎神经干细胞能在体外适宜的培养条件下进行长期的培养、传代及冻存、复苏,并具有多向分化潜能。  相似文献   

11.
Spontaneous intracerebral hemorrhage (ICH) is often a fatal event. In a patient who survives the initial ictus, the resulting hematoma within brain parenchyma can trigger a series of events that lead to secondary insults and severe neurological deficits. Great efforts have been focused on searching for new approaches to help patients recover neurological function after ICH. Previous studies indicate that mesenchymal stem cells (MSCs) grafted into the ischemic rat brain can improve neurological function. However, there is no report regarding whether MSCs can be used in the same way to improve the neurological function after ICH. We generated the ICH model by injecting collagenase VII into rat brain. Subsequently, 5-bromo-2-deoxyuridine (BrdU)-labeled mesenchymal stem cells were delivered into the brain through carotid artery, cervical vein or lateral ventricle. The distribution and differentiation of MSCs were investigated by methods of immunohistochemistry. We found that MSCs were able to differentiate into neural cells in vitro as well as in the rat brain after ICH. The injected MSCs were able to migrate into hippocampus, blooding foci and ipsilateral cortex. In the hippocampus, MSCs differentiated into neurons; but in surrounding bleeding foci, they differentiated into neurons and astrocytes. In the ipsilateral cortex, MSCs differentiated into neurons, astrocytes and oligodendrocytes. Notably, the motor function of the rats in the carotid artery (CA) group and the lateral ventricle (LV) group improved significantly. Collectively, our study indicates that MSCs are able to differentiate into neural cells in the rat brain after ICH and can significantly improve motor function.  相似文献   

12.
In the present study, we attempted to explore cell transplantation therapy for intracerebral hemorrhage (ICH) using embryonic stem (ES) cells. Collagenase-induced ICH rats were used as model animals. Mouse ES cells were differentiated into nestin-positive neural stem cells in vitro by alltrans retinoic acid (ATRA). ATRA-treated ES cells (10(5)) were transplanted into the lateral ventricle in the hemisphere contralateral to the hemorrhage 7 days after collagenase infusion. Twenty-eight days after transplantation, ES-derived neurons and astrocytes were observed around the hematoma cavities of the brain in all of the ten rats receiving grafts. Graft-derived neurons were found in the subependymal area of the lateral ventricle as cellular nodules. Although one of the ten rats receiving grafts showed uncontrolled growth of astroglia derived from the ES cells, intraventricular transplantation of ATRA-treated ES cells is an effective delivery system of neuronal lineage-committed progenitor cells toward the site of ICH.  相似文献   

13.
目前用于脑出血治疗研究的主要细胞有:神经干细胞、遗传工程神经干细胞、骨髓间质干细胞、脐血细胞、胚胎干细胞、重组细胞、微囊化人工细胞等,本文就前5种细胞移植在出血性脑损伤中的保护作用进行综述。干细胞移植在脑出血性动物身上的实验所取得的良好效果,显示细胞移植重建损伤的脑组织、改善脑功能成为治疗脑出血疾病的必然途径。临床实验也取得了一定效果,但要真正应用于临床目前还有许多问题要解决:要想准确的定向诱导细胞分化,需要更深入的研究局部微环境,细胞因子及基因对干细胞分化的作用;细胞移植促进脑出血功能的恢复,其确切的机制还需要进一步研究 ;诱导分化的神经干细胞是否能象正常神经细胞一样能分泌神经递质,是否具有复杂的电生理等。  相似文献   

14.
The present study investigated the ability for adult rat neural progenitor cells to survive transplantation, structurally repopulate the striatum and improve motor function in the quinolinic acid (QA) lesion rat model of Huntington's disease. Neural progenitor cells were isolated from the subventricular zone of adult Wistar rats, propagated in culture and labeled with BrdU (50 microM). Fourteen days following QA lesioning, one group of rats (n = 12) received a unilateral injection of adult neural progenitor cells ( approximately 180,000 cells total) in the lesioned striatum, while a second group of rats (n = 10) received a unilateral injection of vehicle only (sham transplant). At the time of transplantation adult neural progenitor cells were phenotypically immature, as demonstrated by SOX2 immunocytochemistry. Eight weeks following transplantation, approximately 12% of BrdU-labeled cells had survived and migrated extensively throughout the lesioned striatum. Double-label immunocytochemical analysis demonstrated that transplanted BrdU-labeled progenitor cells differentiated into either astrocytes, as visualized by GFAP immunocytochemistry, or mature neurons, demonstrated with NeuN. A proportion of BrdU-labeled cells also expressed DARPP-32 and GAD67, specific markers for striatal medium spiny projection neurons and interneurons. Rats transplanted with adult neural progenitor cells also demonstrated a significant reduction in motor function impairment as determined by apomorphine-induced rotational asymmetry and spontaneous exploratory forelimb use when compared to sham transplanted animals. These results demonstrate that adult neural progenitor cells survive transplantation, undergo neuronal differentiation with a proportion of newly generated cells expressing markers characteristic of striatal neurons and reduce functional impairment in the QA lesion model of Huntington's disease.  相似文献   

15.
16.
Transplantation of human neural stem cells into the dentate gyrus or ventricle of rodents has been reportedly to enhance neurogenesis. In this study, we examined endogenous stem cell proliferation and angiogenesis in the ischemic rat brain after the transplantation of human neural stem cells. Focal cerebral ischemia in the rat brain was induced by middle cerebral artery occlusion. Human neural stem cells were transplanted into the subventricular zone. The behavioral performance of human neural stem cells-treated ischemic rats was significantly improved and cerebral infarct volumes were reduced compared to those in untreated animals. Numerous transplanted human neural stem cells were alive and preferentially localized to the ipsilateral ischemic hemisphere. Furthermore, 5-bromo-2′-deoxyuridine-labeled endogenous neural stem cells were observed in the subventricular zone and hippocampus, where they differentiated into cells immunoreactive for the neural markers doublecortin, neuronal nuclear antigen Neu N, and astrocyte marker glial fibrillary acidic protein in human neural stem cells-treated rats, but not in the untreated ischemic animals. The number of 5-bromo-2′-deoxyuridine-positive ? anti-von Willebrand factor-positive proliferating endothelial cells was higher in the ischemic boundary zone of human neural stem cells-treated rats than in controls. Finally, transplantation of human neural stem cells in the brains of rats with focal cerebral ischemia promoted the proliferation of endogenous neural stem cells and their differentiation into mature neural-like cells, and enhanced angiogenesis. This study provides valuable insights into the effect of human neural stem cell transplantation on focal cerebral ischemia, which can be applied to the development of an effective therapy for stroke.  相似文献   

17.
神经干细胞立体定向脑内移植治疗大鼠重型颅脑损伤   总被引:4,自引:0,他引:4  
目的探讨神经干细胞立体定向脑内移植对颅脑损伤大鼠的治疗效果。方法采用Feeney等人的方法制成大鼠脑损伤模型,伤后24h将体外培养的胚胎神经干细胞经立体定向移植到脑损伤灶内。伤前24h、伤后24h及伤后1、2周行动物神经学缺损评分。结果移植后1和2周,接受神经干细胞移植的大鼠神经学缺损评分明显低于对照组(P〈0.05);且其脑组织切片中的神经元数量较对照组明显增多(P〈0.01)。结论经立体定向移植到脑内损伤灶的神经干细胞可存活、增殖、分化、并可明显改善重型颅脑损伤后大鼠的神经学功能。  相似文献   

18.
目的探讨神经干细胞移植治疗颞叶癫痫的疗效。方法首先建立大鼠的海人酸颞叶癫痫模型。然后从胚胎大鼠的海马中分离培养出神经干细胞,将预诱导的、向氨基丁酸能神经元方向分化的胚胎神经干细胞移植入颞叶癫痫模型的海马内,在移植后1w、2w、3w和4w分别进行移植癫痫大鼠的癫痫最低阂值的测定、海马中氨基丁酸的测定和移植的神经干细胞在体内分化情况的鉴定。结果移植后的癫痫最低阈值和海马中的氨基丁酸逐渐增加,接近但未达到正常值,预诱导的神经干细胞全部分化为氨基丁酸能神经元。结论神经干细胞移植治疗颞叶癫痫具有一定效果。  相似文献   

19.
目的 探讨神经干细胞(NSCs)移植入脑出血模型大鼠脑内后对脑组织内IL-6、TNF-α表达的影响.方法 体外培养大鼠NSCs;选取SD大鼠180只,随机取60只大鼠为健康对照组(A组),余120只采用Ⅳ型胶原酶诱导法建立脑出血模型,并随机分成两组:B组60只为单纯脑出血组;C组60只为脑出血+NSCs移植组,三组大鼠分别在NSCs移植后6 h、1 d、3 d、7 d、14 d进行肢体运动功能评分后,处死取脑组织匀浆,以EHSA方法分别检测三组大鼠脑组织内5个时间点IL-6、TNF-a的表达,并用统计学的方法分析.结果 与单纯脑出血组比较,脑出血+NSCs移植组能够显著降低IL-6、TNF-a表达(P<0.05)和促进肢体功能恢复(P<0.05).结论 大鼠NSCs脑内移植能有效降低脑出血模型大鼠脑内IL-6、TNF-a的表达,并能够改善运动神经功能缺损.  相似文献   

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