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1.
背景:目前针对肝癌是起源于成熟肝细胞的去分化还是肝干细胞的成熟受阻这一问题仍存在争议。 目的:观察胚胎肝干细胞癌变的可能性。 设计、时间和地点:随机对照动物实验,于2006-01/12在中山大学动物实验中心完成。 材料:6~8周龄BALB/c雌性小鼠70只,体质量20~25 g,随机分为3组:空白对照组10只、模型组30只、实验组30只。另取孕14 d胎鼠用于制备胚胎肝细胞。 方法:3组小鼠麻醉后均行2/3肝切除。模型组诱导肝癌,在饮水中加入100 μg/L二乙基亚硝胺,连续饮用12周。实验组在诱导肝癌的基础上行肝干细胞移植,即分离的胚胎肝细胞经肠系膜静脉注射到肝脏,每只小鼠移植1×106个细胞。空白对照组仅行肝干细胞移植,同时饮正常水。6个月后处死小鼠,制备肝脏标本,取肝癌结节做连续切片进行指标检测。 主要观察指标:采用免疫组化或免疫荧光方法检测Y染色体性别决定区域蛋白、甲胎蛋白、胎盘型谷胱苷肽转移酶或细胞角蛋白19的表达,以分析肝癌的细胞来源和特征。 结果:6个月后,实验组有8只小鼠成功诱癌,其中7只为肝细胞性肝癌,1只为胆管细胞性肝癌;模型组有7只小鼠成功诱癌,其中6只为肝细胞性肝癌,1只为胆管细胞性肝癌;两组诱癌率、肿瘤病理类型比较均无明显差异(P > 0.05)。实验组17.1%的肝细胞癌结节Y染色体性别决定区域蛋白染色阳性,胆管细胞癌结节Y染色体性别决定区域蛋白染色均为阴性。肝细胞癌结节表达甲胎蛋白和谷胱苷肽转移酶,而不表达细胞角蛋白19;胆管细胞癌结节不表达甲胎蛋白和谷胱苷肽转移酶,而表达角蛋白19。 结论:在诱导肝癌过程中移植的肝干细胞有癌变的潜能,并保持了胚胎肝干细胞未成熟的特征,仍表达谷胱苷肽转移酶和甲胎蛋白。  相似文献   

2.
背景:目前研究认为表皮干细胞数量及增殖分化潜能的差异,可能是创面愈合能力随年龄增大而逐渐降低的重要原因之一。 目的:探讨不同发育阶段人皮肤表皮干细胞β1整合素、角蛋白 19、p63转录因子和端粒酶反转录酶的表达特征。 设计、时间及地点:细胞学体外对照观察,于2005-09/2007-04在南昌大学第一附属医院烧伤研究所完成。 材料:流产胎儿皮肤标本由南昌大学第一附属医院产科提供,4~12周岁少儿、25~45岁成人烧伤整形植皮手术剩余皮片标本由南昌大学第一附属医院烧伤科提供。 方法:标本取材后立即用中性甲醛固定,常规石蜡包埋,4 ℃保存。所有标本经系列切片后,分别采用鼠抗人角蛋白19、β1整合素、p63转录因子、端粒酶反转录酶单克隆抗体作为一抗,以免疫组织化学EnVision法检测皮肤组织中表皮干细胞标志物的表达。 主要观察指标:200倍光镜下观察免疫组化染色结果,测定表皮干细胞标志物阳性表达率。 结果:胎儿皮肤、少儿皮肤和成人皮肤表皮的基底部、毛囊周围细胞β1整合素、角蛋白19、p63和端粒酶反转录酶均呈棕黄色阳性表达。胎儿表皮基底层细胞β1整合素、角蛋白19和p63均呈强阳性表达;少儿表皮基底层细胞中大部分表达β1整合素、角蛋白19和p63,阳性细胞均匀分布;成人表皮基底层细胞β1整合素、角蛋白19和p63呈弱阳性表达,阳性细胞散在分布;胎儿表皮基底层端粒酶反转录酶阳性细胞表达强度>少儿表皮>成人表皮。随年龄的增加,β1整合素、角蛋白19、p63和端粒酶反转录酶在胎儿、少儿、成人皮肤组织中的阳性表达率呈下降趋势(F=5.06,P < 0.01)。 结论:胎儿皮肤、少儿皮肤和成人皮肤表皮干细胞主要位于表皮基底层,端粒酶表达阳性区域及信号强度与表皮干细胞的定位分布及阳性表达强度相一致。提示表皮干细胞的数量、端粒酶活性表达的强弱可能与不同发育阶段皮肤组织的修复能力差异密切相关。  相似文献   

3.
背景:现阶段对于肝干细胞在胚胎肝发育过程中的形态学特征、时空分布、分化过程的报道很少。 目的:以甲胎蛋白、细胞角蛋白19、c-Met作为标志物,观察其在不同发育阶段肝干细胞中的时空分布。 设计、时间及地点:细胞学体外观察,于2005-06/2006-12在潍坊医学院中心实验室完成。 材料:40例胚胎标本取自3个月内的流产胎儿,由潍坊医学院附属医院提供。 方法:收集流产胚胎,30 min内制作切片,显微镜下根据胚层形成情况、体节数目及器官发育状况确定胎龄。选择胎龄第3~12周的标本切片,每隔10张抽出1张切片,行免疫组织化学染色。 主要观察指标:第3~12周胚胎肝干细胞甲胎蛋白、c-Met、细胞角蛋白19的表达。 结果:第3~5周甲胎蛋白、c-Met呈阳性反应,提示为肝干细胞;第10~12周甲胎蛋白、c-Met阳性细胞主要分布于汇管区周围,提示此时肝干细胞主要局限于汇管区周围的肝索内,与成年肝中卵圆细胞(成年肝干细胞)的分布一致。细胞角蛋白19阳性反应在第7周时开始出现;第10~11周时细胞角蛋白19阳性反应主要位于汇管区附近的肝索细胞和胆管板细胞及胆管上皮细胞;第12周时细胞角蛋白19阳性信号仅见于胆管板和胆管上皮细胞。此时所有的胆管板细胞及胆管上皮细胞均呈甲胎蛋白、c-Met和细胞角蛋白19阳性。 结论:细胞角蛋白19在肝干细胞中无表达,仅在胆管上皮细胞及其祖细胞中有表达,可能不适合作为肝干细胞的标志物。所有的胆管板及胆管上皮细胞均呈甲胎蛋白、c-Met、细胞角蛋白19阳性反应,推测甲胎蛋白+/c-Met+/细胞角蛋白19+的细胞可能为胆管祖细胞。  相似文献   

4.
背景:体外构建组织工程化人工涎腺需获得分化增殖良好、数量充足的种子细胞,但正常下颌下腺中很难分离出成体干细胞。 目的:利用腺体组织损伤动物模型体外分离下颌下腺干/祖细胞,并进行克隆化培养。 设计、时间和地点:细胞学体外观察,于2006-03/2007-01在遵义医学院贵州省细胞工程实验室完成。 材料:8周龄雄性SD大鼠10只,由解放军第三军医大学动物中心提供。 方法:10只大鼠采用结扎下颌下腺主导管、抑制腺体分泌来建立组织损伤模型,1周后切取腺体组织,酶消化法体外分离下颌下腺干/祖细胞,原代培养10~14 d后,挑取培养皿中形成的小的类圆形、类上皮细胞集落予以纯化,传代后进行单克隆培养。 主要观察指标:下颌下腺干/祖细胞免疫细胞化学染色及免疫荧光染色结果,通过绘制生长曲线分析下颌下腺干/祖细胞体外增殖能力。 结果:实验获得表达层粘蛋白的细胞具有干细胞特征,CD29呈阳性表达表明其具有高黏附、高增殖等组织干细胞特性,角蛋白19的阳性表达提示下颌下腺干/祖细胞呈上皮源性。其生长曲线近似“S”形,体外培养增殖活跃。 结论:实验结果显示,下颌下腺干/祖细胞具有组织干细胞的特征,有望成为组织工程化人工涎腺构建的一类种子细胞来源。  相似文献   

5.
背景:胎肝细胞可能具有比骨髓干细胞等更强的增殖分化能力和更低的免疫原性,但目前涉及胎肝干细胞直接分离、培养的报道甚少。 目的:拟在体外分离培养小鼠胎肝干细胞,并对其生物学特性进行初步鉴定。 设计、时间及地点:细胞学体外观察,于2008-03/06在重庆市神经病学重点实验室完成。 材料:SPF级13.5 d龄昆明种胎鼠9只,由重庆医科大学实验动物中心提供。 方法:采用胶原酶+EDTA联合消化法与差速贴壁法体外分离胎鼠肝干细胞,按2×108 L-1接种,待细胞80%~90%汇合后消化传代。采用链霉亲和素-生物素-过氧化酶复合物技术对原代接种后5 d的贴壁细胞进行多种肝干细胞表面标志物的标记。 主要观察指标:原代胎肝干细胞形态变化,胎肝干细胞的传代扩增情况,胎肝干细胞表面标志的表达。 结果:原代培养24 h细胞贴壁,呈致密圆形,边缘清楚;3 d左右部分细胞呈梭形,7 d后细胞铺展呈上皮样;传代后细胞扩增速度无明显变化,至第5代仍保持较均一的上皮细胞状。原代接种后5 d的贴壁细胞,人干细胞因子受体与甲胎蛋白呈阳性表达,白蛋白与细胞角蛋白19呈阴性。 结论:胎肝干细胞原代培养早期表达甲胎蛋白与人干细胞因子受体,不表达白蛋白和细胞角蛋白19,提示所分离的胎肝干细胞可能是一种较原始的干细胞,尚处在未分化的早期阶段。  相似文献   

6.
背景:目前肝干细胞尚无特异性标志物,故其分离、培养尤其是纯化技术尚不成熟。 目的:观察体外培养的小鼠胚胎肝干细胞生物学特性,及其向肝样细胞的诱导分化能力。 设计、时间及地点:细胞学体外观察,于2008-01/06在厦门大学附属中山医院消化中心实验室完成。 材料:SPF级BALB/c胎鼠20只,13.5 d龄,由厦门大学生命科学学院实验动物中心提供。 方法:无菌操作取出胎鼠肝脏,用细胞刮梳理后用IV型胶原酶消化。取分离纯化后的第2代细胞进行免疫荧光染色。细胞培养到第3代时,分别用3 mmol/L丁酸钠盐与0.1% DMSO进行诱导,5 d后收集细胞进行Western blotting实验。 主要观察指标:胚胎肝干细胞的形态及表面分子的表达,诱导后胚胎肝干细胞mRNA与蛋白水平的变化。 结果:分离培养的细胞第2代即可得以纯化,呈克隆样生长,高表达白蛋白、甲胎蛋白、C-met及角蛋白19,且多数细胞共表达白蛋白与角蛋白19、C-met与角蛋白19,双阳性率约80%。细胞诱导后肝细胞标志物白蛋白表达明显增加,而作为不成熟肝细胞的经典标志物甲胎蛋白表达减少,同时干细胞标志物 c-kit mRNA水平也明显下降,角蛋白19水平无明显变化。 结论:胚胎肝干细胞具有双向分化潜能,经丁酸钠盐与DMSO诱导后可以向肝样细胞分化。  相似文献   

7.
背景:多数学者认为正常乳腺组织中无神经内分泌细胞,乳腺病变后出现神经内分泌细胞是乳腺上皮干细胞分化过程中受局部微环境和激素水平影响所发生的突变、异常分化结果。 目的:通过检测人乳腺病变组织中乳腺癌耐药蛋白、细胞角蛋白8及嗜铬蛋白A的表达,从干细胞多向分化的角度探讨人乳腺病变中出现神经内分泌细胞的可能机制。 方法:用乳腺癌耐药蛋白作为SP干细胞标记物,用细胞角蛋白8作为腺上皮分化标记物,用嗜铬蛋白A作为神经内分泌分化指标标记物,采用免疫组化方法分别检测这3种蛋白在89例人乳腺组织中的表达情况,并分析3者间的相关性。 结果与结论:乳腺癌耐药蛋白与细胞角蛋白8在正常乳腺组织、乳腺增生组织、乳腺病变组织中均有表达,乳腺癌耐药蛋白在病变组织中的表达呈上升趋势,细胞角蛋白8的表达则随乳腺组织异常分化程度的降低呈逐渐减少,嗜铬蛋白A只在乳腺病变组织中有表达。在正常乳腺组织、乳腺增生组织和乳腺病变组织中,乳腺癌耐药蛋白与嗜铬蛋白A的阳性表达呈显著相关(P < 0.01),与细胞角蛋白8的阳性表达无明显相关(P=0.069)。上述结果表明正常及增生乳腺组织中未发现神经内分泌细胞,乳腺病变组织中发现神经内分泌细胞,其机制可能与多向分化潜能干细胞的分化有关。  相似文献   

8.
背景:目前肝纤维化尚没有公认的特效疗法,近年来骨髓间质干细胞应用于肝纤维化的治疗已取得一定效果,但具体机制仍不明确。 目的:体外探讨人骨髓间质干细胞对肝星状细胞的调控作用。 设计、时间及地点:细胞学体外观察,于2008-06/12在中山大学干细胞与组织工程研究中心、中山大学第三附属医院中心实验室完成。 材料:骨髓来源于青年健康志愿者髂骨,人肝星状细胞购自中山大学实验动物中心,人正常肝细胞系L-O2购自中山大学实验动物中心。 方法:将分离提纯的人骨髓间质干细胞与肝星状细胞在6孔Transwell板上建立上下共培养体系,接种密度均为2×104cells/well。另设L-O2代替人骨髓间质干细胞作为阴性对照,单纯培养的肝星状细胞为空白对照,培养72 h。 主要观察指标:肝星状细胞形态及免疫细胞化学染色结果,流式细胞仪检测肝星状细胞凋亡情况,Western blot法检测肝星状细胞活化标志α-肌动蛋白的表达。 结果:倒置显微镜下活化的肝星状细胞呈扁平状,胞浆内缺乏脂肪滴,α-肌动蛋白位于肝星状细胞胞质内,呈高张力纤维状分布。与L-02+肝星状细胞组、单纯肝星状细胞组比较,骨髓间质干细胞+肝星状细胞共培养组的肝星状细胞凋亡率明显升高(P < 0.05),α-肌动蛋白表达水平明显下调。 结论:人骨髓间质干细胞可抑制肝星状细胞活化,促进其凋亡,这可能是骨髓间质干细胞抗肝纤维化的作用机制。  相似文献   

9.
肝移植治疗肝原发性鳞状细胞癌1例   总被引:1,自引:0,他引:1  
景:肝原发性鳞状细胞癌较罕见,各种治疗方法疗效不佳,预后差。 目的:随访观察1例肝移植治疗肝原发性鳞状细胞癌患者的近期疗效。 方法:观察中山大学附属第一医院1例肝原发性鳞状细胞癌患者经肝移植治疗后的情况,并回顾性分析迄今为止见于文献报道的国内外65例肝原发性鳞状细胞癌的临床表现、组织来源、治疗方法及预后。 结果与结论:该患者经肝移植后肝功能恢复正常,肿瘤标志物癌胚抗原、糖类抗原199、鳞状细胞癌抗原等均降至正常水平,恢复良好。国内外研究表明,肝原发性鳞状细胞癌的临床表现不典型,组织来源多样,手术切除、化疗等治疗方法疗效不佳,预后差。本例结果显示,肝移植治疗肝原发性鳞状细胞癌近期疗效良好。  相似文献   

10.
背景:动物实验已经证实骨髓间充质干细胞体外诱导可分化为表皮细胞。 目的:观察体外培养条件下人骨髓间充质干细胞向表皮细胞的分化及表皮细胞角蛋白表达。 方法:采用Ficoll-Paque密度梯度离心法提取人胚胎骨髓间充质干细胞,以免疫细胞化学及流式细胞仪测定细胞表面CD33、CD34标记物进行鉴定。取第3代骨髓间充质干细胞以30%条件培养基诱导其向表皮细胞分化,免疫细胞化学染色观察诱导后细胞形态与细胞角蛋白水平变化。 结果与结论:采用密度梯度离心法从人胚胎骨髓中分离培养得到细胞成分均一的骨髓间充质干细胞。骨髓间充质干细胞经体外诱导后,出现细胞角蛋白19表达阳性细胞,说明骨髓间充质干细胞在体外诱导后可能发生向表皮细胞分化。  相似文献   

11.
肝干细胞的分离培养和诱导分化及应用   总被引:1,自引:0,他引:1  
王璐  刘军 《中国神经再生研究》2008,12(12):2355-2358
摘要:近年来,肝干细胞的基础研究已取得重大进展,肝干细胞可分化为肝细胞和胆管细胞已经得到了共识。目前多用两步胶原酶灌流消化法、密度梯度离心法、离心淘洗技术、荧光激活细胞筛选法及免疫磁珠细胞筛选法进行肝干细胞的分离纯化。然而肝干细胞的定向分化是一个复杂的过程,是在一些细胞内外因素及肝脏微环境的作用下完成的。肝脏微环境有利于干细胞的生长、分化与功能发挥。肝细胞生长因子能够促进肝干细胞生长增殖,并且促进其向成熟肝细胞分化,肝细胞转录因子在肝干细胞的分化调控中也起重要作用。肝癌可能是肝干细胞分化不全或分化异常所致,肝干细胞可望作为靶向基因治疗肝癌极具潜力的载体细胞。  相似文献   

12.
背景:角膜缘干细胞体外培养的关键在于建立稳定的体外培养体系,包括角膜缘干细胞的定位、培养条件、载体选择和鉴别方法等。 目的:探索兔角膜缘上皮干细胞体外扩增方法,并对其生物学特性进行鉴定。 方法:采用兔角膜缘组织块培养法,以人羊膜为载体,在体外进行兔角膜缘上皮干细胞原代和传代培养。倒置显微镜观察其体外生长特征;苏木精-伊红染色以及扫描电镜观察其形态;AE5和P63单克隆抗体免疫组织化学染色鉴定其蛋白表达。 结果与结论:采用组织块培养法可在体外获得角膜缘上皮干细胞,能成功传代培养且保持较高增殖潜能。培养于去上皮羊膜上的干细胞可融合成片,呈“拉网”现象。原代角膜缘上皮干细胞AE5单克隆抗体染色阳性率低于5%,P63染色阳性达90%;随传代次数增加AE5染色阳性率增高,P63染色阳性率降低。结果显示兔角膜缘组织块培养法可以在体外成功获得角膜缘上皮干细胞,原代和传代细胞均具有干细胞特性,以羊膜为载体培养可形成角膜移植片。  相似文献   

13.
INTRODUCTION: Von Willebrand factor (VWF) plays a critical role in hemostasis by carrying factor VIII (FVIII) and binding to specific ligands on the surface of blood platelets and within the blood vessel wall. MATERIALS AND METHODS: We constructed a gene-specific phage display library containing small, random VWF fragments. Using this library, we mapped the repertoire of immune epitopes recognized by a commonly used commercial rabbit antihuman VWF polyclonal antibody. RESULTS: A total of eight discrete epitopes within the VWF protein were identified, including two dominant epitopes that account for 74% of immuno selected VWF fragments. Comparison with previously mapped epitopes for mouse monoclonal antibodies reveals four overlapping regions that may identify common antigenic determinants. The distribution of these epitopes was not readily predicted from primary amino acid sequence divergence among these mammalian species or standard algorithms for the prediction of antigenicity, hydrophobicity, or surface probability. CONCLUSION: Taken together with previous monoclonal antibody epitope mapping studies, our results suggest that a limited number of exposed domains on the surface of the human VWF protein may be the primary determinants of immunogenicity.  相似文献   

14.
BACKGROUND: Hepatic veno-occlusive disease (VOD) is one of the most disastrous complications after allogeneic hematopoetic stem cell transplantation (HSCT). Thrombocytopenia with refractoriness to platelet transfusions suggests an increased platelet consumption in these patients. Interactions between platelets and endothelial cells might contribute to the hypercoagulable state at the sinusoidal endothelium as a central mechanism in the pathogenesis of VOD. STUDY DESIGN: The influence of activated platelets on cultured human endothelial cells was investigated in vitro. We focused on the release of plasminogen activator inhibitor-1 (PAI-1) from endothelial cells which has earlier been found to be significantly elevated in plasma of VOD patients. Endothelial cells isolated from human umbilical cords (HUVEC) were incubated with activated platelets. The release of PAI-1 in the presence or absence of specific antibodies was determined by ELISA technique. Tissue factor (TF) expression on endothelial cells was observed by flowcytometric analysis. RESULTS: HUVEC incubated with activated platelets were found to release significantly more PAI-1 compared to untreated cultures. The endothelial PAI-1-secretion after incubation of HUVEC with activated platelets was completely inhibited by an IgG monoclonal antibody against human transforming growth factor beta-1 (TGF beta-1). In contrast, PAI-1 production was not suppressed after inhibition of HUVEC-platelet-interaction by an IgG monoclonal antibody against CD154 (CD40L) expressed on the surface of activated platelets. An increased release of PAI-1 and an increased expression of tissue factor (TF) on the endothelial cell surface were observed after stimulation with TGF beta-1. CONCLUSION: TGF beta-1 released from activated platelets contributes to the hemostatic imbalance at the sinusoidal endothelium in patients with hepatic VOD by increase of endothelial cell PAI-1 production and TF expression. As a potent profibrotic cytokine, TGF beta-1 might further be involved in phlebosclerosis and sinusoidal fibrosis occurring in VOD.  相似文献   

15.
A monoclonal antibody, named BSP-2, has been produced against glycoproteins extracted from neonatal mouse brain. Its reactivity with live cells established the surface location of the antigen. In primary cultures of dissociated cerebellar cells, the antibody bound to neuronal cell types, but not to astrocytes nor to fibroblasts. Immunoprecipitates prepared with the BSP-2 antibody contained a triplet of high-molecular weight glycoproteins with apparent molecular weights of 180,000, 140,000 and 120,000.  相似文献   

16.
背景:E-选择素作为一种细胞黏附分子在细胞间选择性识别黏附、调节白细胞归巢及渗出和肿瘤细胞转移等方面的作用已有研究报道,但胚肝发育过程中E-选择素的定性定位研究、与胚肝造血功能的关系至今未见报道。 目的:观察小鼠肝脏发育过程中E-选择素的表达与肝细胞、肝血窦内皮形态分化及胎肝造血作用的关系。 方法:取胚胎11.5(E11.5)至出生后15.5 d(P15.5)的小鼠胚胎或胎肝,常规制作石蜡切片,行苏木精-伊红染色及免疫组织化学检测。光学显微镜下观察胎肝的组织结构变化及细胞形态;免疫组织化学方法检测发育各期肝组织内E-选择素表达。 结果与结论:E11.5肝始祖细胞形成团索样结构,其间有窦样间隙,内有散在的造血干细胞;E12.5 d肝始祖细胞开始增殖分化,其后,随着小鼠胎肝的不断发育,肝细胞数量及体积增大,核质比减小,至出生时形成肝小叶结构;窦样间隙由少到多,由宽变窄,窦壁内皮细胞从少量、不连续逐步增殖形成完整的内皮;造血细胞于E12.5开始造血,E13.5~E15.5达高峰,之后造血功能均逐步减弱。E-选择素表达于E11.5~E15.5胎肝的内皮细胞,定位于内皮细胞的胞膜,随内皮细胞发育及肝细胞分化成熟,E-选择素表达渐消失。提示E12.5~E15.5为小鼠胎肝各细胞发育分化的关键时期,E-选择素表达于此阶段的血窦内皮,与肝脏造血及肝细胞分化有密切关系。  相似文献   

17.
BACKGROUND: It has been previously shown that hyperbaric oxygen may promote proliferation of neural stem cells and reduce death of endogenous neural stem cells (NSCs).
OBJECTIVE: To explore the effects of hyperbaric oxygen on the differentiation of hypoxic/ischemic brain-derived NSCs into neuron-like cells and compare with high-concentration oxygen and high pressure.
DESIGN, TIME AND SETTING: An in vitro contrast study, performed at Laboratory of Neurology, Central South University between January and May 2006.
MATERIALS: A hyperbaric oxygen chamber (YLC 0.5/1A) was provided by Wuhan Shipping Design Research Institute; mouse anti-rat microtubule-associated protein 2 monoclonal antibody by Jingmei Company, Beijing; mouse anti-rat glial fibrillary acidic protein monoclonal antibody by Neo Markers, USA; mouse anti-rat galactocerebroside monoclonal antibody by Santa Cruz Biotechnology Inc., USA; and goat anti-mouse fluorescein isothiocyanate-labeled secondary antibody by Wuhan Boster Bioengineering Co., Ltd., China.
METHODS: Brain-derived NSCs isolated from brain tissues of neonatal Sprague Dawley rats were cloned and passaged, and assigned into five groups: normal control, model, high-concentration oxygen, high pressure, and hyperbaric oxygen groups. Cells in the four groups, excluding the normal control group, were incubated in serum-containing DMEM/F12 culture medium. Hypoxic/ischemic models of NSCs were established in an incubator comprising 93% N2, 5% 002, and 2% 02. Thereafter, cells were continuously cultured as follows: compressed air (0.2 MPa, 1 hour, once a day) in the high pressure group, compressed air + a minimum of 80% 02 in the hyperbaric oxygen group, and a minimum of 80% Q2 in the high-concentration oxygen group. Cells in the normal control and model groups were cultured as normal.
MAIN OUTCOME MEASURES: At day 7 after culture, glial fibrillary acidic protein, microtubule-associated protein 2, and galactocerebroside immunofluorescence staining were examined to observe differentiation and calculate the percentage of NSCs differentiating into neuron-like cells or neuroglia-like cells.
RESULTS: Neuron-like cells or neuroglia-like cells were visualized in all five groups. There were no significant differences in the percentage of differentiating cells between the hyperbaric oxygen group and the normal control group (P 〉 0.05). The percentage of NSCs differentiating into neuron-like cells in the hyperbaric oxygen group was significantly greater than model, high-concentration oxygen, and high pressure groups; however, the percentage differentiating into neuroglia-like cells was significantly lower (P 〈 0.01 ).
CONCLUSION: Hyperbaric oxygen promotes the differentiation of brain-derived neural stem cells into neuron-like cells but inhibits differentiation into neuroglia-like cells. Furthermore, the efficacy of hyperbaric oxygen is superior to high-concentration oxygen and high pressure.  相似文献   

18.
自体骨髓单个核细胞肝内移植治疗失代偿期肝硬化   总被引:1,自引:0,他引:1  
背景:大量研究发现,骨髓间充质干细胞是肝干细胞的主要肝外来源,骨髓间充质干细胞在特定环境下可分化为肝组织和肝细胞从而参加肝脏的修复与重建。 目的:观察自体骨髓单个核细胞移植治疗失代偿期肝硬化的效果及安全性。 方法:选择肝硬化失代偿期患者20例,在无菌条件下由髂后上棘抽取骨髓200 mL,在体外分离纯化骨髓单个核细胞并制成10 mL细胞悬液,经肝动脉将细胞悬液移植入肝脏,分别在移植后第2、4、8、12周复查肝脏功能,观察实验室指标,临床症状及不良反应。 结果与结论:移植后2周内所有血清学指标均无显著改变,移植后4周白蛋白由(27.05±5.23) g/L升到(30.02±5.02) g/L,纤维蛋白原由(1.55±0.53) g/L升到(2.55±0.53) g/L,凝血酶原时间由(24.05±5.23) s降到 (17.05±5.13) s,胆碱酯酶和甲胎蛋白也有不同程度上升,氨基转移酶、总胆红素水平在移植前后无显著性变化。单个核细胞移植后食欲改善、体力好转10例,腹胀减轻6例,腹水减少7例,下肢浮肿减轻4例。全部患者在移植中未发生严重并发症,移植近期无不良反应出现。  相似文献   

19.
BACKGROUND: It has been previously shown that hyperbaric oxygen may promote proliferation of neural stem cells and reduce death of endogenous neural stem cells (NSCs).OBJECTIVE: To explore the effects of hyperbaric oxygen on the differentiation of hypoxic/ischemic brain-derived NSCs into neuron-like cells and compare with high-concentration oxygen and high pressure.DESIGN, TIME AND SETTING: An in vitro contrast study, performed at Laboratory of Neurology,Central South University between January and May 2006.MATERIALS: A hyperbaric oxygen chamber (YLC 0.5/1A) was provided by Wuhan Shipping Design Research Institute; mouse anti-rat microtubute-associated protein 2 monoclonal antibody by Jingmei Company, Beijing; mouse anti-rat glial fibrillary acidic protein monoclonal antibody by Neo Markers,USA; mouse anti-rat galactocerebroside monoclonal antibody by Santa Cruz Biotechnology Inc.,USA; and goat anti-mouse fluorescein isothiocyanate-labeled secondary antibody by Wuhan Boster Bioengineering Co., Ltd., China.METHODS: Brain-derived NSCs isolated from brain tissues of neonatal Sprague Dawiey rats werecloned and passaged, and assigned into five groups: normal control, model, high-concentration oxygen, high pressure, and hyperbaric oxygen groups. Cells in the four groups, excluding the normal control group, were incubated in serum-containing DMEM/F12 culture medium. Hypoxic/ischemic models of NSCs were established in an incubator comprising 93% N2, 5% CO2, and 2% O2.Thereafter, cells were continuously cultured as follows: compressed air (0.2 MPa, 1 hour, once a day)in the high pressure group, compressed air+a minimum of 80% O2 in the hyperbaric oxygen group,and a minimum of 80% O2 in the high-concentration oxygen group. Cells in the normal control and model groups were cultured as normal.MAIN OUTCOME MEASURES: At day 7 after culture, glial fibrillary acidic protein,microtubule-associated protein 2, and galactocerebroside immunofluorescence staining were examined to observe differentiation and calculate the percentage of NSCs differentiating into neuron-like cells or neuroglia-like cells.RESULTS: Neuron-like cells or neuroglia-like cells were visualized in all five groups. There were no significant differences in the percentage of differentiating cells between the hyperbaric oxygen group and the normal control group (P>0.05). The percentage of NSCs differentiating into neuron-like cells in the hyperbaric oxygen group was significantly greater than model, high-concentration oxygen, and high pressure groups; however, the percentage differentiating into neureglia-like cells was significantly lower (P<0.01).CONCLUSION: Hyperbaric oxygen promotes the differentiation of brain-derived neural stem cells into neuron-like cells but inhibits differentiation into neuroglia-like cells. Furthermore, the efficacy of hyperbaric oxygen is superior to high-concentration oxygen and high pressure.  相似文献   

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