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神经干细胞(neural stem cells,NSCs)是具有高度自我更新能力,能分化为神经元及胶质细胞的前体细胞.NSCs增殖及分化调控机制的研究对于其治疗神经系统退行性疾病及功能修复具有重大意义.研究表明:神经干细胞的治疗作用包括干细胞的归巢,定植和激活自体神经干细胞,但更重要的是神经干细胞所分泌的神经细胞调控因子群(neurocyte growth regulatory factors,NGRFs)的药理学作用.深入研究这些因子不但对神经细胞再生及功能重建有重要意义,而且对研究这些因子的药理作用机理及开发大分子新药具有指导意义. 相似文献
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目的:探讨胚胎神经干细胞的分离、培养和鉴定方法.方法:原代培养、培养细胞生长状况观察、组织化学鉴定、流式细胞仪进行细胞凋亡分析.结果:准确分离出了神经干细胞,了解其生长规律及细胞凋亡的情况.结论:用此方法分离胚胎神经干细胞实用、便捷和可行. 相似文献
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目的：采用实时荧光定量聚合酶链式反应（polymerase chain reaction,PCR）技术,研究P2X受体在大鼠神经胶质瘤和嗜铬细胞瘤及原代培养皮层神经元和星形胶质细胞上的表达差异。方法：取新生1-2 d SD大鼠大脑皮层,分离纯化神经元和星形胶质细胞,并采用实时荧光定量PCR技术,比较P2X受体在大鼠胶质瘤C6细胞、大鼠肾上腺嗜铬细胞瘤PC-12细胞、星形胶质细胞和皮层神经元上的表达差异。结果：C6细胞P2X2、P2X3和P2X5表达水平显著高于星形胶质细胞,P2X4、P2X6和P2X7表达水平显著低于星形胶质细胞,PC-12细胞P2X1、P2X2、P2X3和P2X6表达水平显著高于皮层神经元,P2X5和P2X7表达水平则显著低于皮层神经元。此外,还发现P2X2、P2X5和P2X6在C6和PC-12细胞上的表达水平存在显著差异。结论：大鼠胶质瘤和嗜铬细胞瘤细胞表达多种P2X受体,且与原代培养细胞存在表达差异,提示核苷酸介导的信号传递系统可能作为潜在的肿瘤治疗靶点。 相似文献
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项目的主要目标是实现猪的诱导多能干细胞(i PSCs)向神经干细胞(NSCs)体外诱导。在整个实验探索的过程中,我们使用了传统的诱导分化形成神经干细胞(NSCs)的方法,即通过拟胚体(EB)的形成并添加维甲酸(RA)进行诱导。猪的诱导多能干细胞(i PSCs)经历了传代培养、拟胚体(EB)时期、诱导分化时期三个重要阶段,生成一类新的细胞。最后我们使用免疫荧光法检测到新的细胞中存在神经干细胞的特异性标志物Nestin、神经元标志物β-tubulin III以及神经胶质细胞标志物GFAP表达,从而确定这类新的细胞为神经干细胞。可见,猪的诱导多能干细胞(i PSCs)经过这种传统诱导分化方法培养可分化为具备基础特征的神经干细胞(NSCs),能够表达出神经干细胞(NSCs)特异的标志分子nestin。 相似文献
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大鼠骨髓间充质干细胞的分离、培养及生物学鉴定 总被引：1，自引：0，他引：1
目的:探讨成体大鼠的间充质干细胞(MSCs)的分离、体外培养,为其应用提供理论依据.方法:用密度梯度离心结合贴壁培养法分离纯化大鼠骨髓MSCs,传代扩增,对分离的细胞进行碱性磷酸酶的检测.结果:取得较高纯度的成体大鼠骨髓MSCs,并保持细胞的活性;成体大鼠骨髓MSCs在体外培养中为贴壁生长的单个核球形细胞,培养3～4d后开始大量增殖,并形成形态均一的细胞增殖集群,对分离后所获得的细胞进行AKP染色为强阳性.结论:采用密度梯度离心结合贴壁培养法可获得较高纯度的MSCs,是实用、便捷和可行的方法.并且在体外培养条件下能大量增殖,形成形态均一的细胞集落,可以成为进一步进行细胞扩增或其他实际应用的基础. 相似文献
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目的:探讨影响大鼠胚胎成纤维细胞(REF)分离和培养的一些因素,以建立稳定的REF培养体系.方法:取SD大鼠胚胎分离成纤维细胞,利用体外培养体系,对REF的生长形态、生长曲线进行观察,以探讨不同胚胎日龄以及不同胰酶作用时间对REF分离及培养的影响,并对其进行免疫细胞化学鉴定.结果:13.5d 胎龄鼠胚的REF分离效果优于10.5d、18.5d 胎龄鼠胚;REF在体外为贴壁生长型细胞,第三代细胞增殖旺盛;并表达波形蛋白(vimentin)、层黏连蛋白(laminin,LN)和纤维连接蛋白(fibronectin,FN)蛋白.结论:13.5d 胎龄SD大鼠REF以H-DMEM做培养基,在第3代增殖旺盛,合成多种细胞外基质,最适宜作胚胎干细胞或其它悬浮培养细胞的饲养层. 相似文献
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神经干细胞是指具有分化为神经元、星形胶质细胞、少突胶质细胞的能力,能进行自我更新并分化形成新的脑组织细胞的细胞.作为干细胞中最特殊的一种,神经干细胞的研究工作已成为近年来脑科学研究的一个重要领域,也必将成为21世纪生命科学研究的热点. 相似文献
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目的:研究人参皂甙Rg1对小鼠脂肪干细胞神经样分化的促进作用,并初步探讨其作用机理。创新点:证明了人参皂甙Rg1可以通过mi RNA-124途径促进脂肪干细胞的神经样分化。方法:从BALB/c小鼠腹股沟和睾丸脂肪垫处分离培养脂肪干细胞,利用流式细胞仪检测分离的脂肪干细胞纯度。试验分为以下五组:磷酸缓冲盐溶液(PBS)组、3-异丁基-1-甲基黄嘌呤(IBMX)组、IBMX+Rg1低剂量组、IBMX+Rg1中剂量组和IBMX+Rg1高剂量组。用细胞免疫组化方法检测了脂肪干细胞向神经样细胞的分化效率,用荧光定量聚合酶链式反应(qP CR)方法检测mi RNA-124的表达变化,用免疫印迹的方法检测巢蛋白(nestin)、βIII-微管蛋白(βIII-tubulin)及羧基端小结构域磷酸酶1(SCP1)的表达水平。结论:免疫组化结果显示,IBMX可以成功诱导小鼠脂肪干细胞向神经样细胞的分化;免疫印迹结果显示,Rg1可以显著提高神经样细胞标记蛋白的表达水平;荧光定量PCR结果显示,Rg1可以促进miRNA-124的表达量,进而降解神经分化抑制因子SCP1的表达,促进脂肪干细胞的神经样分化效率。 相似文献
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1IntroductionNeural stem cells have great potential for use intreating neural damage and neurodegenerative disor-ders such as Parkinson’s Disease and Alzhei mer’s Di-sease.They are undifferentiated elements found inboth the embryonic and adult brain[1].Growth factor-responsive cells fromthe embryonic and adult controlnervous system(CNS)were isolated and culturedinvitrointhe early1990’s[2-4].Thelocation of CNS stemcells in the adult brain were therefore identified[5]mainlyinthe striatum,… 相似文献
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概述了神经干细胞的生物学特性、分布以及影响其分化和增殖的因素，并探讨了神经干细胞移植和基因治疗在神经系统疾病治疗中的应用. 相似文献
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This paper aims to screen and identify sphere clone cells with characteristics similar to cancer stem cells in human gallbladder
cancer cell line GBC-SD. GBC-SD cells were cultured in a serum-free culture medium with different concentrations of the chemotherapeutic
drug cisplatin for generating sphere clones. The mRNA expressions of stem cell-related genes CD133, OCT-4, Nanog, and drug
resistance genes ABCG2 and MDR-1 in sphere clones were detected by quantitative real-time polymerase chain reaction (PCR).
Stem cell markers were also analyzed by flow cytometry and immunofluorescent staining. Different amounts of sphere clones
were injected into nude mice to test their abilities to form tumors. Sphere clones were formed in serum-free culture medium
containing cisplatin (30 μmol/L). Flow cytometry results demonstrated that the sphere clones expressed high levels of stem
cell markers CD133+ (97.6%) and CD44+ (77.9%) and low levels of CD24+ (2.3%). These clones also overexpressed the drug resistance genes ABCG2 and MDR-1. Quantitative real-time PCR showed that
sphere clones expressed stem cell genes Nanog and OCT-4 284 and 266 times, respectively, more than those in the original GBC-SD
cells. Immunofluorescent staining showed that sphere clones overexpressed OCT-4, Nanog, and SOX-2, and low expressed MUC1
and vimentin. Tumor formation experiments showed that 1×103 sphere clone cells could induce much larger tumors in nude mice than 1×105 GBC-SD cells. In conclusion, sphere clones of gallbladder cancer with stem cell-like characteristics can be obtained using
suspension cultures of GBC-SD cells in serum-free culture medium containing cisplatin. 相似文献
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绞股蓝皂甙和大豆皂甙对神经干细胞分化的影响 总被引：1，自引：0，他引：1
Neural stem cell has a potential to differentiate into neurons, astrocytes and oligodendrocytes. It provides an in vitro model to screen herbal medicines on the cellular differentiation and development level. In this work, active component from gypenosides and soyasaponins was prepared to investigate their effects on the differentiation of neural stem cells.. Both gypenosides and soyasaponins promote the differentiation of neural stem cells. This method provides speed and practicality for screening effective herbal medicine. It is well suited for studying the mechanism of cell differentiation and development. 相似文献
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Fu Y Wang SQ Liu YP Wang GP Wang JT Gong SS 《Journal of Zhejiang University. Science. B》2008,9(4):299-305
Objective: To evaluate the transduction efficiency of a recombinant adenovirus carrying the gene for green fluorescent protein (Ad-GFP) into the primary cultures of fetal neural stem cells (NSCs) by the expression of GFP. Methods: The Ad-GFP was constructed by homologous recombination in bacteria with the AdEasy system; NSCs were isolated from rat fetal hippocampus and cultured as neurosphere suspensions. After infection with the recombinant Ad-GFP, NSCs were examined with a fluorescent microscopy and a flow cytometry for their expression of GFP. Results: After the viral infection, flow cytometry analysis revealed that the percentage of GFP-positive cells was as high as 97.05%. The infected NSCs sustained the GFP expression for above 4 weeks. After differentiated into astrocytes or neurons, they continued to express GFP efficiently. Conclusion: We have successfully constructed a viral vector Ad-GFP that can efficiently infect the primary NSCs. The reporter gene was showed fully and sustained expression in the infected cells as well as their differentiated progenies. 相似文献
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目的体外分离胰腺干细胞,选择最佳培养条件探索胰腺干细胞在体外环境下向β细胞的分化情况.方法取SD大鼠胰腺组织,胶原酶消化,密度梯度离心获得纯化的胰腺导管上皮细胞.采用分步诱导法诱导胰腺导管上皮细胞向胰岛β细胞分化;用胰岛素释放实验检测胰岛功能,免疫荧光法检测nestin,PDX-1,CK-19,CK-20胰岛素及胰高血糖素等的表达.结果胰腺消化培养6～12h,可看到胰腺导管上皮细胞贴壁生长,通过4步诱导培养,nestin阳性细胞快速生长,并可分泌胰岛素.结论体外分离的成人胰岛前体细胞,在诱导因子的作用下,胰腺干细胞可定向分化为β细胞,有望应用于糖尿病的治疗. 相似文献
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探讨体外条件下骨骼肌卫星细胞的原代培养、纯化及鉴定的方法。取新生3～5 d的SD大鼠处死清洗后,采用Ⅰ型胶原酶、胰蛋白酶联合酶消化30 min,细胞悬液过滤离心后,用含有10%胎牛血清的培养液悬浮细胞,1 h后重新接种,重新接种2次,最后接种于L-多聚赖氨酸铺底的培养瓶中;采用骨骼肌肌动蛋白(α-sarcometric actin)免疫荧光方法鉴定骨骼肌细胞。肌卫星细胞培养5～6 h后开始贴壁,48 h完全贴壁,之后细胞进一步增多并相互融合,逐渐按一定方向呈有序排列。当细胞增殖至70%密度时,有融合趋势,加入分化培养基培养48 h后可形成多核的肌管,行免疫荧光染色,90%以上的细胞α-sarcometric actin染色阳性,证明培养的为骨骼肌细胞。 相似文献
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This article presents the findings of a quasi-experimental study that evaluated the effect of differentiated instruction on students’ learning, in mixed ability classrooms. Participants in the study were 24 teachers and 479 grade-four elementary students. Results indicate that in classrooms where differentiated instruction methods were systematically employed, students made better progress compared to students in classrooms where differentiated instruction methods were not employed, the family's socioeconomic status did not lead to differentiation in students’ achievement and the quality of differentiated teaching had a corresponding effect on students’ achievement. Based on these findings, the article discusses the significance of the systematic employment of differentiated instruction methods in mixed ability classrooms for promoting equity, optimization of quality and effectiveness in teaching. 相似文献