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Flk1^+间充质干细胞减轻四氯化碳导致的肝纤维化的研究 被引量:9
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作者 史明霞 房佰俊 +3 位作者 廖联明 杨少光 刘煜昊 赵春华 《生物工程学报》 CAS CSCD 北大核心 2005年第3期396-401,共6页
许多慢性肝脏疾病都会发生肝纤维化,但是目前尚缺乏对肝纤维化切实有效的治疗手段。实验发现,Flk1(fetalliverkinase)阳性间充质干细胞(MSC)能够减轻四氯化碳(CCl4 )所致小鼠肝纤维化。取雄性BALB c小鼠骨髓,分离培养Flk1+ MSC ,用CCl4... 许多慢性肝脏疾病都会发生肝纤维化,但是目前尚缺乏对肝纤维化切实有效的治疗手段。实验发现,Flk1(fetalliverkinase)阳性间充质干细胞(MSC)能够减轻四氯化碳(CCl4 )所致小鼠肝纤维化。取雄性BALB c小鼠骨髓,分离培养Flk1+ MSC ,用CCl4 制作雌性小鼠肝纤维化模型,在CCl4 损伤后立即或1周后经尾静脉注射Flk1+ MSC ,2或5周后检测受体小鼠肝脏的纤维化程度和供体细胞的植入。结果发现,CCl4 损伤后立即注射Flk1+ MSC ,可以使肝脏损伤程度明显减轻,减少胶原沉积,使肝脏羟脯氨酸含量及血清纤维化指标显著下降;而损伤1周后注射细胞则无明显变化。免疫荧光、PCR和荧光原位杂交方法证实,在受体肝脏中有供体细胞植入,呈上皮细胞形态,并表达白蛋白,但是数量很少。因此,Flk1+ MSC具有潜在的植入肝组织的能力,并可能启动肝组织的内源性修复,减轻CCl4 导致的肝纤维化。 展开更多
关键词 充质细胞 纤维化 移植
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体外诱导小鼠胎肝间充质细胞向神经元样细胞分化的研究 被引量:4
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作者 刘平平 张洹 刘革修 《第二军医大学学报》 CAS CSCD 北大核心 2003年第9期950-953,共4页
目的 :探讨胎肝间充质细胞在体外向神经元样细胞分化的潜能。 方法 :无菌条件下获取胎龄 1 4 .5d的胎肝细胞 ,DMEM/HEPES/F1 2 (2 0 %FCS)培养得到贴壁细胞并传代 ,第 5代细胞经 β 巯基乙醇 (β mercaptoethanol,β ME)与全反式维甲酸 ... 目的 :探讨胎肝间充质细胞在体外向神经元样细胞分化的潜能。 方法 :无菌条件下获取胎龄 1 4 .5d的胎肝细胞 ,DMEM/HEPES/F1 2 (2 0 %FCS)培养得到贴壁细胞并传代 ,第 5代细胞经 β 巯基乙醇 (β mercaptoethanol,β ME)与全反式维甲酸 (all trans rentinoicacid ,ATRA)程序化诱导处理后 5h、5d ,用免疫细胞化学鉴定神经元抗原表达 ,同时用半定量RT PCR检测神经元相关基因mRNA表达情况。 结果 :胎肝间充质细胞经 β ME、ATRA诱导处理后 ,约 80 %细胞呈现典型的神经元样的形态表型。免疫细胞化学显示诱导分化后的细胞表达神经元特异蛋白如神经元核特异性抗原 (neuron specificnuclearpro tein ,NeuN)、神经元特异性管蛋白Ⅲ型 (neuronc specifictubulinⅢ ,TuJ 1 )、神经丝蛋白中链 (neurofilament M ,NF M )及神经元特异性烯醇化酶 (neuron specificenolase ,NSE) ,但是无成熟神经元微管相关蛋白Tau(microtubule associatedproteintau ,Tau 1 )、微管相关蛋白 2 (microtubule associatedprotein 2 ,MAP 2 )表达。半定量RT PCR结果显示诱导分化后的细胞酪氨酸羟化酶 (tyrosinehydroxylase ,TH)mRNA有表达 ,脑因子 1基因 (brainfactor 1 ,BF 1 )、脑因子 3基因 (brain 3,Brn 3)、神经丝蛋白轻链基因 展开更多
关键词 体外诱导 小鼠 充质细胞 神经元样细胞 细胞分化
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TAT-PDX1蛋白诱导的人胎肝间充质干细胞治疗NOD/SCID鼠糖尿病 被引量:2
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作者 姜铧 张洹 +2 位作者 李东升 梁清乐 李雪峰 《南京医科大学学报(自然科学版)》 CAS CSCD 北大核心 2009年第9期1199-1203,共5页
目的:观察人胎肝间充质干细胞在TAT-PDX1蛋白的作用下体外分化为胰岛β细胞的能力以及治疗NOD/SCID鼠糖尿病的效果。方法:体外分离纯化获得人胎肝间充质干细胞,用TAT-PDX1蛋白体外诱导分化为胰岛β细胞,制作NOD/SCID鼠糖尿病模型,并将... 目的:观察人胎肝间充质干细胞在TAT-PDX1蛋白的作用下体外分化为胰岛β细胞的能力以及治疗NOD/SCID鼠糖尿病的效果。方法:体外分离纯化获得人胎肝间充质干细胞,用TAT-PDX1蛋白体外诱导分化为胰岛β细胞,制作NOD/SCID鼠糖尿病模型,并将诱导后的细胞移植到糖尿病鼠肾包膜下,观察各组小鼠血糖变化并行IPGTT试验。结果:用TAT-PDX1蛋白体外诱导分化的细胞稳定表达胰岛素,移植到糖尿病鼠肾包膜下,能快速降低血糖水平,取出移植物,血糖水平再次升高。结论:TAT-PDX1蛋白诱导的人胎肝间充质干细胞能有效降低糖尿病鼠血糖,有望成为糖尿病细胞治疗的种子细胞。 展开更多
关键词 人胎充质细胞 TAT-PDX1蛋白 糖尿病模型
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TAT-PDX1融合蛋白诱导人胎肝间充质干细胞向胰岛β细胞分化 被引量:1
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作者 姜铧 张洹 +1 位作者 李东升 梁清乐 《中国组织工程研究与临床康复》 CAS CSCD 北大核心 2009年第1期48-52,共5页
背景:PDX1是决定胰岛β细胞功能的关键因子,应用穿膜蛋白TAT将PDX1蛋白导入细胞内,若能发挥诱导分化作用将使干细胞治疗糖尿病向临床应用更进一步。目的:观察TAT-PDX1融合蛋白诱导人胎肝间充质干细胞向胰岛β细胞分化的能力。设计、... 背景:PDX1是决定胰岛β细胞功能的关键因子,应用穿膜蛋白TAT将PDX1蛋白导入细胞内,若能发挥诱导分化作用将使干细胞治疗糖尿病向临床应用更进一步。目的:观察TAT-PDX1融合蛋白诱导人胎肝间充质干细胞向胰岛β细胞分化的能力。设计、时间及地点:细胞学体外观察,于2007-05/2008-03在郧阳医学院附属太和医院湖北省胚胎干细胞研究重点实验室完成。材料:流产胎儿来源于肝功能正常、HBsAg(-)的健康产妇,TAT-PDX1融合蛋白由美国佛罗里达大学医学院唐东启博士惠赠。方法:采用贴壁法体外分离培养人胎肝间充质干细胞,传至第3代加入20mol/LTAT-PDX1融合蛋白诱导,分别于第4,6,8,10天向诱导细胞中加入含葡萄糖的DMEM/F12培养基。主要观察指标:诱导后细胞形态变化及双硫腙染色结果,RT-PCR检测胰岛β细胞特异基因的表达,放射免疫法测定胰岛素累积分泌量。结果:人胎肝间充质干细胞易于体外分离和扩增,加入TAT-PDX1融合蛋白诱导后细胞由梭形逐渐变圆,并形成胰岛样细胞团,双硫腙染色呈棕红色。诱导第4,6,8,10天细胞序贯表达胰岛β细胞特异基因,NeuroD基因最先出现,随后消失,Nkx2.2,Pax4,Nkx6.1,ISL-1基因随后依次出现。诱导后胰岛素累积分泌量逐渐增加,葡萄糖刺激后2-4周细胞胰岛素基础分泌量和刺激后胰岛素分泌量均显著增加,且可以维持在较高水平。结论:TAT-PDX1融合蛋白能诱导人胎肝间充质干细胞向胰岛β细胞分化。 展开更多
关键词 人胎充质细胞 TAT-PDX1蛋白 胰岛Β细胞 分化
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骨髓间充质干细胞对大鼠急性肝损伤修复的影响 被引量:6
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作者 梁昌宇 覃山羽 +3 位作者 姜海行 王东旭 苏思标 梁梓宇 《世界华人消化杂志》 CAS 北大核心 2009年第12期1178-1184,共7页
目的:观察骨髓间充质干细胞(BMSCs)移植通过抑制RhoA-ROCK信号转导通路对大鼠急性肝损伤修复过程的影响.方法:贴壁筛选法培养、纯化♂SD大鼠BMSCs.将健康♀SD大鼠随机分为3组:正常对照组(N组,n=10)、CCl4组(C组,n=10)及CCl4+BMSCs组(T组... 目的:观察骨髓间充质干细胞(BMSCs)移植通过抑制RhoA-ROCK信号转导通路对大鼠急性肝损伤修复过程的影响.方法:贴壁筛选法培养、纯化♂SD大鼠BMSCs.将健康♀SD大鼠随机分为3组:正常对照组(N组,n=10)、CCl4组(C组,n=10)及CCl4+BMSCs组(T组,n=10).N组不予任何处理;T组和C组腹腔注射0.1mL/100g600mL/LCCl4花生油溶液制造急性肝损伤模型后24h,分别经鼠尾静脉移植的间充质干细胞和等量PBS.各组于不同时间点留取标本,采用HE染色和血清肝功能酶学指标观察受损肝脏恢复过程;RT-PCR方法检测RhoA mRNA的表达;Western blot检测RhoA蛋白的表达.结果:与C组相比,T组移植BMSCs后能显著改善CCl4急性损伤大鼠肝功能(1d,ALT:89.70±3.09U/Lvs147.59±6.83U/L,AST:263.67±17.05U/Lvs472.68±19.04U/L,P<0.01或0.05;7d,ALT:42.38±14.31U/Lvs92.75±6.70U/L,AST173.85±16.80U/Lvs260.41±25.35U/L,均P<0.05),并迅速修复肝脏结构.N组大鼠肝脏RhoA mRNA和蛋白表达量极低,C组经CCl4损伤后RhoA mRNA和蛋白表达量迅速增加(1.39±0.046vs0.57±0.010,1.23±0.020vs0.35±0.036,均P<0.01),此后表达量缓慢降低.T组经BMSCs移植后,与C组比较,RhoA mRNA和蛋白表达水平迅速下降.结论:Rho-ROCK信号转导通路参与CCl4导致的急性肝损伤发生、发展和修复全过程.BMSCs可能通过抑制RhoA-ROCK信号转导通路加速受损肝脏修复. 展开更多
关键词 急性损伤:骨髓充质细胞 RHOA Rho—ROCK信号转导通路
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人胎肝间充质干细胞体外分离培养和生物学鉴定 被引量:3
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作者 何念海 赵文利 王宇明 《肝脏》 2005年第3期182-185,共4页
目的建立人胎儿肝脏非实质细胞间充质干细胞(nonparenchymalmesenchymalstemcells,NPMSC)的分离、培养方法,观察NPMSC形态学、细胞生物学特性及细胞表型特征,以获得大量人源性间充质干细胞(MSC)。方法采用体外细胞培养技术分离培养人胎... 目的建立人胎儿肝脏非实质细胞间充质干细胞(nonparenchymalmesenchymalstemcells,NPMSC)的分离、培养方法,观察NPMSC形态学、细胞生物学特性及细胞表型特征,以获得大量人源性间充质干细胞(MSC)。方法采用体外细胞培养技术分离培养人胎儿NPMSC,用显微摄像、MTT和图像分析研究其增殖及生长特征,用流式细胞仪和免疫组化法鉴定其表型。结果每个胎儿肝脏可获得10000±2000个贴壁细胞,可见5~10个高速增生的细胞集落。原代和传代培养均生长缓慢,按1∶3传代,传1代需8~10d,细胞经传代后逐渐纯化,传10代后可达109个细胞。流式细胞仪检测NPMSC不表达CD34,而表达CD166。对传代NPMSC进行人甲胎蛋白和白蛋白免疫组化分析,细胞表达微量甲胎蛋白,不表达白蛋白。结论肝非实质细胞中含有MSC,且量较大,可成为种子细胞。NPMSC在普通培养条件下不向肝细胞分化。 展开更多
关键词 人胎 充质细胞 体外培养 分离培养 细胞培养 生物学鉴定
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同时扩增与收获脐带血来源造血干细胞及间充质干细胞的研究 被引量:5
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作者 范秀波 宋克东 +4 位作者 刘天庆 殷轶群 郝永杰 马学虎 崔占峰 《高校化学工程学报》 EI CAS CSCD 北大核心 2010年第5期808-818,共11页
目的为利用生物反应器,实现脐带血(Umbilical cord blood,UCB)来源的造血干细胞(Hematopoietic stem cells,HSCs)与间充质干细胞(Mesenchymal stem cells,MSCs)的同时扩增与收获。方法为在不添加血清、只添加细胞因子组合(SCF15ng·... 目的为利用生物反应器,实现脐带血(Umbilical cord blood,UCB)来源的造血干细胞(Hematopoietic stem cells,HSCs)与间充质干细胞(Mesenchymal stem cells,MSCs)的同时扩增与收获。方法为在不添加血清、只添加细胞因子组合(SCF15ng·mL-1,FL5ng·mL-1,TPO6ng·mL-1,IL-315ng·mL-1,G-CSF1ng·mL-1,GM-CSF5ng·mL-1)及海藻酸钙壳聚糖胶珠包被基质细胞支持的条件下,采用玻璃包被的聚苯乙烯(Glass coated styrene copolymer,GCSC)微载体与生物反应器相结合的策略,考察了UCB-HSCs与UCB-MSCs在转瓶及旋转壁式生物反应器(Rotating wall vessel bioreactor,RWVB)内的共培养。结果RWVB中的扩增效果最佳,12天内有核细胞(Nuclear cells,NCs)扩增了3.7±0.3倍;集落形成细胞(Colony-forming units in culture,CFU-Cs)扩增了5.1±1.2倍;CD34+CD45+CD105-(HSCs)细胞扩增了5.2±0.4倍;CD34-CD45-CD105+(MSCs)细胞扩增了13.9±1.2倍。培养结束后,通过自由沉降的方法分离UCB-HSCs和粘附在GCSC微载体表面的UCB-MSCs。同时,细胞多向诱导分化及免疫表型分析结果显示,粘附在GCSC微载体表面上的细胞能够向骨、软骨及脂肪细胞分化;并能够表达间质细胞相关表面标志CD13,CD44,CD73和CD105,而不表达造血细胞的相关表面标志CD34,CD45及HLA-DR,与骨髓MSCs相一致。结论为添加细胞因子、基质细胞及微载体支持的条件下,在生物反应器内能够实现UCB-HSCs和UCB-MSCs的同时扩增与收获。 展开更多
关键词 脐带血 造血干细胞 间充质肝细胞 生物反应器 扩增 收获
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肝衰竭的细胞治疗:距离临床有多远? 被引量:3
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作者 段学章 辛绍杰 《传染病信息》 2011年第3期132-135,共4页
肝衰竭患者因发病急、病死率高常须肝移植。除此之外,可能替代肝移植的细胞治疗是目前研究的热点。本文概述了近年来肝细胞移植、造血干细胞移植、间充质干细胞移植的基础研究进展和临床应用现况,及肝前体细胞和诱导性多能干细胞的特点... 肝衰竭患者因发病急、病死率高常须肝移植。除此之外,可能替代肝移植的细胞治疗是目前研究的热点。本文概述了近年来肝细胞移植、造血干细胞移植、间充质干细胞移植的基础研究进展和临床应用现况,及肝前体细胞和诱导性多能干细胞的特点和临床应用前景。 展开更多
关键词 功能衰竭 细胞移植 造血干细胞 间充质肝细胞 诱导性多能干细胞
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骨髓间充质干细胞移植对急性肝功能衰竭大鼠血清及肝组织HMGB1表达的影响 被引量:2
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作者 郑盛 杨涓 +3 位作者 张帆 王玉波 杨晋辉 唐映梅 《中华器官移植杂志》 CAS CSCD 2015年第12期714-719,共6页
目的 探讨骨髓间充质干细胞(BMSC)移植对急性肝功能衰竭(ALF)大鼠血清及肝组织高迁移率族蛋白B1 (HMGB1)表达的影响.方法 将健康雌性SD大鼠用随机数字表法分为空白对照组、ALF组和BMSC移植组,其中ALF组腹腔注射D-氨基半乳糖(900 ... 目的 探讨骨髓间充质干细胞(BMSC)移植对急性肝功能衰竭(ALF)大鼠血清及肝组织高迁移率族蛋白B1 (HMGB1)表达的影响.方法 将健康雌性SD大鼠用随机数字表法分为空白对照组、ALF组和BMSC移植组,其中ALF组腹腔注射D-氨基半乳糖(900 mg/kg)+脂多糖(10μg/kg)建立ALF模型;BMSC移植组腹腔注射D-氨基半乳糖(900mg/kg)+脂多糖(10 μg/kg)后2h,经尾静脉注射BMSC(1.0×10^7);同时空白对照组腹腔注射9 g/L氯化钠溶液1 ml.三组大鼠均在移植后12、24、72 h时处死6只,取血液及肝组织标本,检测大鼠血清丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST),酶联免疫吸附试验法检测血清HMGB1水平,HE染色观察肝组织病理变化,免疫组织化学法检测肝组织HMGB1的表达情况,逆转录聚合酶链反应法检测肝组织HMGB1mRNA.各组间指标差异采用方差分析,同时观察每组大鼠的24 h存活率,并用卡方检验比较其差异.结果 实验大鼠成功诱导为ALF模型.ALF组大鼠血清ALT和AST水平随时间推移逐渐升高.移植后72 h,BMSC移植组的血清ALT和AST与ALF组相比较,差异有统计学意义(均P<0.01).空白对照组血清HMGB1浓度、肝组织HMGB1 mRNA表达以及肝组织HMGB1蛋白表达在各时间点均较低,ALF组及BMSC移植组上述指标均在较高水平,ALF组随时间延长而升高,BMSC移植组移植后随时间延长逐渐下降;在移植后24和72 h时,BMSC移植组相应指标与ALF组相比较,差异有统计学意义(均P<0.01).移植后24 h,空白对照组、ALF组、BMSC移植组大鼠死亡率分别为0(0/24)、33.3%(8/24)和16.7%(4/24),差异有统计学意义(x2=21.098,P<0.01).结论 BMSC移植可以改善ALF大鼠的肝功能以及肝脏病理改变,同时降低ALF大鼠血清及肝组织中HMGB1的表达. 展开更多
关键词 间充质肝细胞 骨髓 功能衰竭 急性 高迁移率族蛋白1 大鼠
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骨髓来源的间充质干细胞对胃癌细胞SGC-7901增殖的影响 被引量:1
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作者 唐辉蓉 崔进 +4 位作者 廖陈 李为明 张家骅 吴雪松 代佑果 《中华肿瘤防治杂志》 CAS 北大核心 2016年第S2期17-18,共2页
目的探讨骨髓来源的间充质肝细胞对胃癌细胞CGC-7901增殖的影响,为胃癌治疗提供新的思路和价值依据。方法通过MTT比色法对传代培养胃癌细胞SGC-7901、5-Fu、间充质肝细胞及原代培养骨髓来源的间充质肝细胞对胃癌细胞SGC-79-1的影响惊醒... 目的探讨骨髓来源的间充质肝细胞对胃癌细胞CGC-7901增殖的影响,为胃癌治疗提供新的思路和价值依据。方法通过MTT比色法对传代培养胃癌细胞SGC-7901、5-Fu、间充质肝细胞及原代培养骨髓来源的间充质肝细胞对胃癌细胞SGC-79-1的影响惊醒分析。结果 1d内MSC组胃癌细胞SGC-7901吸光度比其他两组明显降低,差异显著,P<0.05;MCN组与DMEM组相比,1d后胃癌细胞SGC-7901吸光度也明显下降,但3d时两组差异不明显,无统计学意义(P>0.05)。结论骨髓来源的间充质肝细胞系统5-Fu对胃癌细胞SGC-7901增值抑制作用明显,为胃癌治疗提供了新思路,可知道胃癌的临床治疗。 展开更多
关键词 骨髓来源 间充质肝细胞 胃癌细胞SGC-7901 影响
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适合鸭瘟病毒弱毒增殖的MHC单倍型鸭胚肝间充质干细胞的建立
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作者 佟相慧 问婉欣 +1 位作者 陈洪岩 韩凌霞 《中国兽医科学》 CAS CSCD 北大核心 2020年第2期183-188,共6页
为了建立适于鸭瘟病毒(DPV)弱毒增殖的细胞系,利用4个主要组织相容性复合体(MHC)单倍型鸭品系,体外分离鸭胚肝细胞,在干细胞因子、白细胞抑制因子和成纤维细胞生长因子诱导下,获得鸭胚肝间充质干细胞(DuLMSC)。接种DPV疫苗株C-KCE后,结... 为了建立适于鸭瘟病毒(DPV)弱毒增殖的细胞系,利用4个主要组织相容性复合体(MHC)单倍型鸭品系,体外分离鸭胚肝细胞,在干细胞因子、白细胞抑制因子和成纤维细胞生长因子诱导下,获得鸭胚肝间充质干细胞(DuLMSC)。接种DPV疫苗株C-KCE后,结果显示:F1~F10代均能稳定出现CPE;将F10病毒接种细胞24 h后,通过扫描电镜在胞质、胞核及细胞间隙均能够观察到典型的疱疹病毒粒子;F1至F10代均能扩增出DPV UL2基因片段,序列测定结果表明F1、F5、F10代的扩增产物与NCBI中公布的DPV株(EU082088)的核苷酸同源性为100%;组织培养半数感染量和一步生长曲线结果表明,源自B4系单倍型鸭的DuLMSC细胞系增殖DPV的病毒含量明显高于B1、B2和B3系。本研究为稳定培养鸭瘟病毒提供了优良稳定的实验材料。 展开更多
关键词 鸭瘟病毒 鸭胚充质细胞 主要组织相容性复合体单倍型
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Bone marrow-derived mesenchymal stem cells protect against experimental liver fibrosis in rats 被引量:71
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作者 Dong-ChangZhao Jun-XiaLei +4 位作者 RuiChen Wei-HuaYu Xiu-MingZhang Shu-NongLi PengXiang 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第22期3431-3440,共10页
AIM: Recent reports have shown the capacity of mesenchymal stem cells (MSCs) to differentiate into hepatocytes in vitro and in vivo. MSCs administration could repair injured liver, lung, or heart through reducing infl... AIM: Recent reports have shown the capacity of mesenchymal stem cells (MSCs) to differentiate into hepatocytes in vitro and in vivo. MSCs administration could repair injured liver, lung, or heart through reducing inflammation, collagen deposition, and remodeling. These results provide a clue to treatment of liver fibrosis. The aim of this study was to investigate the effect of infusion of bone marrow (BM)-derived MSCs on the experimental liver fibrosis in rats. METHODS: MSCs isolated from BM in male Fischer 344 rats were infused to female Wistar rats induced with carbon tetrachloride (CCI4) or dimethylnitrosamine (DMN). There were two random groups on the 42nd d of CCI4: CCl4/MSCs, to infuse a dose of MSCs alone; CCI4/saline, to infuse the same volume of saline as control. There were another three random groups after exposure to DMN: DMN10/MSCs, to infuse the same dose of MSCs on d 10; DMN10/saline, to infuse the same volume of saline on d 10; DMN20/MSCs, to infuse the same dose of MSCs on d 20. The morphological and behavioral changes of rats were monitored everyday. After 4-6 wk of MSCs administration, all rats were killed and fibrosis index were assessed by histopathology and radioimmunoassay. Smooth muscle alpha-actin (alpha-SMA) of liver were tested by immunohistochemistry and quantified by IBAS 2.5 software. Male rats sex determination region on the Y chromosome (sry) gene were explored by PCR. RESULTS: Compared to controls, infusion of MSCs reduced the mortality rates of incidence in CCl4-induced model (10% vs 20%) and in DMN-induced model (20-40% vs 90%).The amount of collagen deposition and alpha-SMA staining was about 40-50% lower in liver of rats with MSCs than that of rats without MSCs. The similar results were observed in fibrosis index. And the effect of the inhibition of fibrogenesis was greater in DMN10/MSCs than in DMN20/MSCs. The sry gene was positive in the liver of rats with MSCs treatment by PCR. CONCLUSION: MSCs treatment can protect against experimental liver fibrosis in CCMnduced or DMN-induced rats and the mechanisms of the anti-fibrosis by MSCs will be studied further. 展开更多
关键词 Mesenchymal stem cells Liver fibrosis RAT THERAPY
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Rat bone marrow mesenchymal stem cells differentiate into hepatocytes in vitro 被引量:35
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作者 xin-QinKang Wei-JinZang +6 位作者 Tu-ShengSong Xiao-LiXu Xiao-JiangYu Dong-LingLi Ke-WeiMeng Sheng-LiWu Zhi-YingZhao 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第22期3479-3484,共6页
AIM: To investigate the mechanism and regulation of differentiation from bone marrow mesenchymal stem cells (MSCs) into hepatocytes and to find a new source of celltypes for therapies of hepatic diseases. METHODS: MSC... AIM: To investigate the mechanism and regulation of differentiation from bone marrow mesenchymal stem cells (MSCs) into hepatocytes and to find a new source of celltypes for therapies of hepatic diseases. METHODS: MSCs were isolated by combining gradient density centrifugation with plastic adherence. The cells were cultured in osteogenic or adipogenic differentiation medium and determined by histochemical staining. MSCs were plated in plastic culture flasks that were not coated with components of extracellular matrix (ECM). When MSCs reached 70% confluence, they were cultured in low glucose Dulbecco's modified Eagle's medium supplemented with 10 mL/L fetal bovine serum, 20 ng/mL hepatocyte growth factor (HGF) and 10 ng/mL fibroblast growth factor-4 (FGF-4). The medium was changed every 3 d and stored for albumin, alpha-fetoprotein (AFP) and urea assay. Glycogen store of hepatocytes was determined by periodic acid-Schiff staining.RESULTS: By combining gradient density centrifugation with plastic adherence, we isolated a homogeneous population of cells from rat bone marrow and differentiated them into osteocytes and adipocytes. When MSCs were cultured withFGF-4 and HGF, approximately 56.6% of cells became smallround and epithelioid on d 24 by morphology. Compared with the control, levels of AFP increased significantly from d 12 to 15.5±1.4 μg/L (t = 2.31, P<0.05) in MSCs cultured with FGF-4and HGF, and were higher (46.2±1.5 μg/L)ond 21 (t = 41.926, P<0.01), then decreased to 24.8±2.2 μg/L on d 24 (t = 10.345, P<0.01). Albumin increased significantly on d 21 (t= 3.325, P<0.01) to 1.4±0.2 μg/mL,and to 2.1±0.7 μg/mL on d 24 (t= 3.646, P<0.01). Urea(2.3±0.4 mmol/L) was first detected on d 21 (t = 6.739, P<0.01), and continued to increase to 2.6±0.9 mmol/Lon d 24 (t= 4.753, P<0.01). Glycogen storage was first seen on d 21.CONCLUSION: The method combining gradient density centrifugation with plastic adherence can isolate MSCs. Rat MSCs may be differentiated into hepatocytes by FGF-4 and HGF. Cytokines may play a more important role in differentiation from rat MSCs into hepatocytes. 展开更多
关键词 Mesenchymal stem cell DIFFERENTIATION HEPATOCYTE
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Mesenchymal stem cells rescue acute hepatic failure by polarizing M2 macrophages 被引量:3
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作者 yan-wei li chong zhang +3 位作者 qiu-ju sheng han bai yang ding xiao-guang dou 《World Journal of Gastroenterology》 SCIE CAS 2017年第45期7978-7988,共11页
AIM To investigate whether M1 or M2 polarization contributes to the therapeutic effects of mesenchymal stem cells(MSCs) in acute hepatic failure(AHF).METHODS MSCs were transfused into rats with AHF induced by D-galact... AIM To investigate whether M1 or M2 polarization contributes to the therapeutic effects of mesenchymal stem cells(MSCs) in acute hepatic failure(AHF).METHODS MSCs were transfused into rats with AHF induced by D-galactosamine(DGal N). The therapeutic effects of MSCs were evaluated based on survival rate and hepatocyte proliferation and apoptosis. Hepatocyte regeneration capacity was evaluated by the expression of the hepatic progenitor surface marker epithelial cell adhesion molecule(Ep CAM). Macrophage polarization was analyzed by M1 markers [CD68,tumor necrosis factor alpha(TNF-α),interferon-γ(IFN-γ),inducible nitric oxide synthase(INOS)] and M2 markers [CD163,interleukin(IL)-4,IL-10,arginase-1(Arg-1)] in the survival and death groups after MSC transplantation.RESULTS The survival rate in the MSC-treated group was increased compared with the DPBS-treated control group(37.5% vs 10%). MSC treatment protected rats with AHF by reducing apoptotic hepatocytes and promoting hepatocyte regeneration. Immunohistochemical analysis showed that MSC treatment significantly increased the expression of Ep CAM compared with the control groups(P < 0.001). Expression of Ep CAM in the survival group was significantly up-regulated compared with the death group after MSC transplantation(P = 0.003). Transplantation of MSCs significantly improved the expression of CD163 and increased the gene expression of IL-10 and Arg-1 in the survival group. IL-4 concentrations were significantly increased compared to the death group after MSC transplantation(88.51 ± 24.51 pg/m L vs 34.61 ± 6.6 pg/m L,P < 0.001). In contrast,macrophages showed strong expression of CD68,TNF-α,and INOS in the death group. The concentration of IFN-γ was significantly increased compared to the survival group after MSC transplantation(542.11 ± 51.59 pg/m L vs 104.07 ± 42.80 pg/m L,P < 0.001).CONCLUSION M2 polarization contributes to the therapeutic effects of MSCs in AHF by altering levels of anti-inflammatory and pro-inflammatory factors. 展开更多
关键词 Acute hepatic failure Mesenchymal stem cells MACROPHAGES POLARIZATION INFLAMMATION
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Transplanted bone marrow stromal cells are not cellular origin of hepatocellular carcinomas in a mouse model of carcinogenesis 被引量:1
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作者 Jin-Fang Zheng Li-Jian Liang 《World Journal of Gastroenterology》 SCIE CAS CSCD 2008年第19期3015-3020,共6页
AIM:To investigate the malignant potential of hepatic stem cells derived from the bone marrow stromal cells (BMSCs) in a mouse model of chemical hepatocarcino- genesis. METHODS:BMSCs from male BALB/c mice were harvest... AIM:To investigate the malignant potential of hepatic stem cells derived from the bone marrow stromal cells (BMSCs) in a mouse model of chemical hepatocarcino- genesis. METHODS:BMSCs from male BALB/c mice were harvested and cultured, then transplanted into female syngenic BALB/ c mice via portal vein. Hepato-carcinogenesis was induced by 6 mo of treatment with diethylnitrosamine (DEN). Six months later, the liver was removed from each treated mouse and evaluated by immunohistochemistry and fluorescence in situ hybridization (FISH). RESULTS:Twenty-six percent of recipient mice survived and developed multiple hepatocellular carcinomas (HCCs). Immunohistochemically, HCC expressed placental form of glutathione-S-transferase (GST-P) and α-fetoprotein, but did not express cytokeratin 19. Y chromosome positive hepatocytes were detected by fluorescent in situ hybridization (FISH) in the liver of mice treated with DEN after BMSCs transplantation while no such hepatocytes were identified in the liver of mice not treated with DEN. No HCC was positive for the Y chromosome by FISH. CONCLUSION:Hepatic stem cells derived from the bone marrow stromal cells have a low malignant potential in our mouse model of chemical hepatocarcingenesis. 展开更多
关键词 Bone marrow stromal cell Stem cell HEPATOCARCINOGENESIS Animal study
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Current status and future prospects of mesenchymal stem cell therapy for liver fibrosis 被引量:5
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作者 Yang GUO Bo CHEN +2 位作者 Li-jun CHEN Chun-feng ZHANG Charlie XIANG 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2016年第11期831-841,共11页
Liver fibrosis is the end-stage of many chronic liver diseases and is a significant health threat. The only effective therapy is liver transplantation, which still has many problems, including the lack of donor source... Liver fibrosis is the end-stage of many chronic liver diseases and is a significant health threat. The only effective therapy is liver transplantation, which still has many problems, including the lack of donor sources, immu- nological rejection, and high surgery costs, among others. However, the use of cell therapy is becoming more prev- alent, and mesenchymal stem cells (MSCs) seem to be a promising cell type for the treatment of liver fibrosis. MSCs have multiple differentiation abilities, allowing them to migrate directly into injured tissue and differentiate into hepatocyte-like cells. Additionally, MSCs can release various growth factors and cytokines to increase hepatocyte regeneration, regress liver fibrosis, and regulate inflammation and immune responses. In this review, we summarize the current uses of MSC therapies for liver fibrosis and suggest potential future applications. 展开更多
关键词 Liver fibrosis Cell therapy Mesenchymal stem cells
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