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Co-culture with cardiomyocytes induces mesenchymal stem cells to differentiate into cardiomyocyte-like cells and express heart development-associated genes

Co-culture with cardiomyocytes induces mesenchymal stem cell...

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江西省第四次中西医结合心血管学术交流会

作者： Junyi Zeng~1,Yang Wang~2,Yunfeng Wei~1,An Xie~2,Yuanlei Lou~2,Mei Zhang~1 1 Department of Cardiology,2 Institute of Urological Surgery,the First Affiliated Hospital of Nanchang University

＜正＞Recent studies have demonstrated that bone marrow mesenchymal stem cells（MSCs） have the potential to transdifferentiate into cardiomyocytes（CMs） in an appropriate condition both in vivo and in vitro,and our previous studies also confirmed it.However,there exists a controversy among a variety of studies about whether MSCs can differentiate into CMs without physical contacts with CMs,and the molecular signals that underlie this process are not fully understood.In the present study,MSCs isolated from adult rats were cocultured with CMs obtained from neonatal rat ventricles at 1:5 ratio in a dual chamber dish separated by a semipermeable membrane for 2 weeks.During this non-direct contact coculture procedure,cardiomyogenic differentiation and relative genes expression in MSCs were evaluated. After coculture with CMs,MSCs showed a stick-like or elliptical morphology.Immunofluorescent stain results revealed that a -actin and cardiac troponin T（cTnT） positive cells were found in MSCs at 5 days after coculture with CMs,and 29.63%of MSCs were positive forα-actin and 27.38%for cTnT at 14 days. Results from RT-PCR demonstrated the expression level of TGF-β,Nkx-2.5,GATA-4 and MEF-2C genes began to increase at 1 day and reached the peak at 7 days after coculture.In conclusion,non-direct contact with CMs（1:5） is conducive for MSCs to differentiate into CMs in vitro,and TGF-β,Nkx-2.5, GATA-4 and MEF-2C genes may play a crucial role during the transdifferentiation.

关键词： bone marrow mesenchymal stem cells non - direct contact differentiation genes cardiomyocytes

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Role of over-expression of STAT3 in oncological differention ofMSCs in tumor microenviroment

Role of over-expression of STAT3 in oncological differention...

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第五届全国生物信息学与系统生物学学术大会暨国际生物信息...

作者： Xiangrong Cui Lu Bai Jing Zhu Ministry of Education Key Laboratory of Child Development and Disorders Chongqing Medical University

Objective:To determine the role of over-expression and activation of STAT3 and P-STAT3 on oncological differention of bone marrow mesenchymal stem cell(MSCs) in the tumor microenviroment. Methods:Through MSCs respectively with C6 glioma and actrocytes indirect co-culture to construct the microenvironment of MSCs growth.The experiment was divided into four groups,including the experimental group(MSCs and C6 glioma indirect co-culture),positive control group(the conventional cultured C6 glioma cells),negative control group(MSCs and astrocytes indirect co-culture),blank group(routinely cultured MSCs).STAT3,CyclinDl and BCL-xl in each group were measured by RT-QPCR and Western blot.P-STAT3 in each group were measured by Western blot.The proliferation of each goup cells were measured by MTT. The nude mice tumor formation were measured by HE staining. Results:Compared with negative control group and blank group,STAT3,CyclinDl and BCL-xl mRNA and protein and P-STAT3 in experimental group were significantly increased(Pcells were deeply stained,obviously gathered and arranged in irregular and a large number of neovascularization within the tumor. Conclusion:The over expression and activation of STAT3 and P-STAT3 is one of the important reason for the oncological differention of MSCs in tumor microenviroment.

关键词： STAT3 P-STAT3 tumor microenvironment bone marrow mesenchymal stem cells

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间接接触共培养下骨髓间充质干细胞分化为心肌样细胞及相关调控基因的时序表达

间接接触共培养下骨髓间充质干细胞分化为心肌样细胞及相关调控基...

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第一届全国中西医结合心血管病中青年医师论坛

作者：曾俊义魏云峰汪泱谢安娄远蕾张梅应国秋徐信群南昌大学第一附属医院心血管内科南昌大学第一附属医院泌尿外科研究所

目的:应用新生大鼠心肌细胞(cardiomyocytes,CMs)与骨髓间充质干细胞(mesenchymal stem cells,MSCs)体外间接接触共培养的方法模拟心肌内环境,研究模拟环境下 MSCs 向 CMs 的分化情况及相关调控基因的时序表达,筛选 MSCs 定向分化为心... 详细信息

目的:应用新生大鼠心肌细胞(cardiomyocytes,CMs)与骨髓间充质干细胞(mesenchymal stem cells,MSCs)体外间接接触共培养的方法模拟心肌内环境,研究模拟环境下 MSCs 向 CMs 的分化情况及相关调控基因的时序表达,筛选 MSCs 定向分化为心肌细胞的重要调控基因。方法:MSCs 及 CMs 按1:5的比例进行体外间接接触共培养,连续观察两周,相差显微镜下观察 MSCs 形态变化,荧光免疫组化方法鉴定心肌特征性肌动蛋白α(α-actin)和肌钙蛋白(cTnT)的表达,应用半定量 RT-PCR 分析 TGF-β、Nkx-2.5、GATA-4、MEF-2C、TEF-1等相关调控基因在分化过程中的时序表达。结果:共培养前 MSCs 呈成纤维细胞样,共培养后细胞形态发生变化,细胞体积增大, 梭形形态逐渐变短变粗,近似棒状或椭圆形,细胞之间形成连接,排列方向趋于一致。α-actin 及 cTnT 共培养前无表达,共培养后两周时阳性细胞比例分别为29.63%和27.38%。TGF-β、Nkx- 2.5、GATA-4和 MEF-2C 基因在共培养后一天表达开始增强,诱导后七天达高峰,以后虽有所下降但仍维持在较高水平;TEF-1基因在诱导过程中表达无明显变化。结论:间接接触共培养条件下 MSCs 可分化为心肌样细胞;且在此过程中,TGF-β、Nkx-2.5、GATA-4和 MEF-2C 基因可能是调控 MSCs 定向分化为心肌样细胞的重要调控基因。

关键词： bone marrow mesenchymal stem cells Non-direct contact Differentiation Genes Cardiomyocytes

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Activation of Notch1 signaling promotes c-KitPOS BMCSCs differentiation in vitro

Activation of Notch1 signaling promotes c-KitPOS BMCSCs diff...

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第六届海南省生命科学联合学术会议

作者： Junli Guo Xiaofan Jiang Ranran Ding Yanping Ha Zhenliang Wang Tianfa Li Rujia Li Hanguo Jiang Shaojiang Zheng Zhihua Shen Wei Jie Cardiovascular Institute of Affiliated Hospital Hainan Medical College Department of Pathology School of Basic Medicine ScienceGuangdong Medical College

Aims:c-KitPOS heart tissue-derived cardiac stem cells(CSCs)are helpful in myocardial infraction therapy but that the isolation and maintaining of heart tissue-derived CSCs are potential damage and complex.Whether the bone marrow deserves the seed cells alternative for heart tissue-derived CSCs is poorly defined.Notch signaling plays a critical role in the differentiation of stem cells and its role in c-Kitbone marrow-derived cardiac stem cells(c-KitMCSC)are not well established.Methods and Results:After infected the Sprague Dawley rat bone marrow mesenchymal stem cells(BMSCs)with Notchl intracellular domain(NICD)adenovirus expression plasmid(NICD-Ad)for 8 days,the Notchl signaling was activated and cells underwent the myocardiocyte,smooth muscle cell and endothelial cell lineage differentiation.The c-KitBMCSCs were successfully isolated from the BMSCs by using magnetic activated cell sorting(MACS)plus single cell cloning method.Among the four types of Notchl receptors,Notchl was predominantly expressed that tested by flow cytometry.Treated the c-Kit BMCSCs with Jagged 1,the exogenous Notchl ligand,led to an obviously activation of Notchl signaling and thus drove the cells to prominently differentiate into cardiomyocytes while much slight into smooth muscle cells and epithelial cells.However,the Notchl inhibitor DAPT attenuated Jagged1-induced Notchl activation and multi-lineage differentiation.Conclusions:There are c-KitBMCSCs in the pool of BMSCs,suggesting an alternative seed cell for c-Kitheart tissue-derived CSCs.The activation of Notchl signaling contributes to the c-KitBMCSCs multi-lineage differentiation,with prominent differentiation into cardiomyocytes,implying modulation of Notchl signaling may be have potential application in the stem cell translational medicine.

关键词： bone marrow mesenchymal stem cells c-Kit bone marrow-derived cardiac stem cells Notch 1 signaling Cell differentiation

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