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人重组表皮生长因子和牛磺酸体外诱导人脐血干细胞分化为神经元样细胞
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作者 金玮 邢怡桥 +2 位作者 杨安怀 杨燕宁 艾明 《中华眼底病杂志》 CAS CSCD 北大核心 2009年第4期261-264,共4页
目的探讨人重组表皮生长因子(rhEGF)和牛磺酸体外诱导人脐血间充质干细胞(MSC)分化为神经元样细胞的实验条件和可能机制。方法用含有10^5U/L青霉素、100mg/L链霉素、10%胎牛血清(FBS)、5%自体血浆、4mmol/L一谷氨酰胺、30ng... 目的探讨人重组表皮生长因子(rhEGF)和牛磺酸体外诱导人脐血间充质干细胞(MSC)分化为神经元样细胞的实验条件和可能机制。方法用含有10^5U/L青霉素、100mg/L链霉素、10%胎牛血清(FBS)、5%自体血浆、4mmol/L一谷氨酰胺、30ng/mlrhEGF的Dulbecco改良Eagle培养基与F12以1:1混合(DMEM/F-12)培养液对人脐血MSC进行原代培养,传至第3代后改用含牛磺酸的培养基进行诱导。流式细胞仪检测细胞表面抗原CD90、CD29、CD34、CD44及CD45的表达。免疫细胞化学方法观察神经元特异性烯醇化酶(NSE)、视紫红质、巢蛋白的阳性细胞表达情况。统计学方法采用两组资料的卡方检验分析NSE、视察红质、巢蛋白的表达。结果脐血MSC首次培养5~7d,有间充质样细胞贴壁,3~4周后细胞达80%~90%融合,传代后生长加速;形态学类似骨髓源性MSC。脐血MSC均一稳定地表达MSC的相关抗原CD29、CD44、CD90,不表达CD34、CD45。经牛磺酸体外诱导后的脐血MSC中,3120个细胞中有2515个细胞表达神经元特异性烯醇化酶(NSE),3050个细胞中有903个细胞表达巢蛋白,3175个细胞中有1168个细胞表达视紫红质;未诱导组的2965个脐血MSC中仅有234个细胞表达NSE,其他抗原未见表达。两组间NSE、巢蛋白和视紫红质的表达差异具有统计学意义(X2=3242.9,1073.0,1444.5;P值均〈0.01)。结论人脐血MSC经rhEGF和牛磺酸诱导后能使部分MSC向神经元样细胞或视紫红质阳性细胞分化。 展开更多
关键词 细胞/细胞 脐血干细胞 细胞培养技术/方法 细胞分化/免疫学
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体外培养及诱导胚兔视网膜及脑神经干细胞分化的研究
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作者 吴婵 董方田 +1 位作者 陈连凤 许卓再 《中华眼底病杂志》 CAS CSCD 北大核心 2009年第4期265-269,共5页
目的比较体外不同培养条件对视网膜及脑神经干细胞(NSCs)视紫红质的纯化能力及不同诱导条件对NSCs的诱导分化能力。方法取胚兔视网膜及大脑皮质制备单细胞悬液,分别在5种不同培养基中进行体外培养并纯化。选取无血清条件下传3代的NSC... 目的比较体外不同培养条件对视网膜及脑神经干细胞(NSCs)视紫红质的纯化能力及不同诱导条件对NSCs的诱导分化能力。方法取胚兔视网膜及大脑皮质制备单细胞悬液,分别在5种不同培养基中进行体外培养并纯化。选取无血清条件下传3代的NSCs,分别在2种培养基中诱导8~10d。采用免疫荧光法及流式细胞仪检测所获得细胞的神经干细胞和视网膜神经上皮细胞抗原的表达。结果免疫荧光法显示无血清培养条件下2种组织来源的NSCs均部分表达巢蛋白。流式细胞术显示2种诱导方式均部分细胞表达巢蛋白,较诱导前明显降低,而视紫红质及Thyl.1的表达均较诱导前明显增高。经5%胎牛血清(FBS)诱导的细胞表达视紫红质较联合应用全反式视黄酸(ATRA)时高,差异均有统计学意义(X2=30.59,6.76;P〈0.01),但Thyl.1表达低于后者,差异也有统计学意义(X2=6.19,22.92;P=0.01)。结论无血清培养基添加N2、EGF、bFGF、LIF4种因子可以获得最佳纯化效果。2种诱导培养基都能够诱导NSCs的分化,视网膜NSCs分化为视网膜神经上皮细胞的能力高于大脑皮质NSCs。 展开更多
关键词 细胞/细胞 细胞培养技术/方法 细胞分化/免疫学 神经上皮细胞/细胞
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体外诱导大鼠骨髓间充质干细胞向视网膜色素上皮细胞分化的可行性研究
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作者 高斐 董方田 《中华眼底病杂志》 CAS CSCD 北大核心 2009年第4期254-256,共3页
目的探讨体外诱导大鼠骨髓间充质干细胞(rMSCs)向视网膜色素上皮(RPE)细胞分化的可行性。方法采用贴壁筛选法分离、培养Brown—Norway(BN)rMSCs。将反复冻融制成的BN大鼠RPE细胞裂解液加入到rMSCs培养体系中,鉴定被诱导的细胞是... 目的探讨体外诱导大鼠骨髓间充质干细胞(rMSCs)向视网膜色素上皮(RPE)细胞分化的可行性。方法采用贴壁筛选法分离、培养Brown—Norway(BN)rMSCs。将反复冻融制成的BN大鼠RPE细胞裂解液加入到rMSCs培养体系中,鉴定被诱导的细胞是否同时表达RPE细胞的特征性标记物细胞角蛋白(CK)与S一100。结果经RPE细胞裂解液诱导的rMSCs生长速度减慢,细胞呈长梭形,周边有毛刺样突起。免疫印迹法和双重免疫荧光标记显示部分经诱导的细胞同时表达CK与S一100。流式细胞术显示14.1%的细胞能够同时表达CK与S-100。结论rMSCs经RPE细胞裂解液诱导后能够向RPE细胞方向分化。 展开更多
关键词 细胞/细胞 骨髓祖代细胞/免疫 细胞培养技术/方法 细胞分化/免疫学 色素上皮 眼/细胞 动物 实验
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Extremely well-differentiated adenocarcinoma of the stomach: Clinicopathological and immunohistochemical features 被引量:19
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作者 Takashi Yao Takashi Utsunomiya +2 位作者 Masafumi Oya Kenichi Nishiyama Masazumi Tsuneyoshi 《World Journal of Gastroenterology》 SCIE CAS CSCD 2006年第16期2510-2516,共7页
AIM: Minimal deviation carcinoma of the uterine cervix, otherwise known as extremely well-differentiated adenocarcinoma (EWDA), is characterized by its benign microscopic appearance in contrast to its aggressive be... AIM: Minimal deviation carcinoma of the uterine cervix, otherwise known as extremely well-differentiated adenocarcinoma (EWDA), is characterized by its benign microscopic appearance in contrast to its aggressive behavior. In order to elucidate the clinicopathological features and biological behavior of the gastric counterpart of EWDA, we, using immunohistochemistry, analyzed nine lesions for the phenotypic expression, proliferative activity, and the expression of oncogene-associated products. METHODS: Clinicopathological features, including preoperative biopsy diagnosis, were reviewed. Using immunohitstochemistry, Ki-67 labeling index and expression of p53 and c-erbB-2 protein in the gastric lesions were detected.RESULT: Locations in the middle or upper third of the stomach and polypoid macroscopic features are characteristic of EWDA of the stomach. Although 4 of the 9 lesions showed only focal lymphatic or venous invasion, lymph node metastasis was not present and none of the patients died of the lesions (mean follow-up period, 56 too). All 9 cases of EWDA could be classified into gastric phenotype (5 lesions) and intestinal phenotype (4 lesions). The former resembled gastric foveolar epithelium, mucous neck cells or pyloric glands, but their papillary structures were frequently elongated and the tumor cellsand their nuclei were slightly larger and more hyperchromatic compared to normal epithelium. The latter resembled intestinal metaplasia with minimal nulcear atypia and irregular glands; two of these lesions demonstrated complete intestinal phenotype, while two demonstrated incomplete intestinal phenotype. Ki-67 labeling index was low and none of the cases revealed over-expression of p53 and c-erbB-2 protein. CONCLUSION: Unlike minimal deviation carcinoma of the cervix, these findings suggest that EWDA of the stomach is a lesion of low-grade malignancy. This favorable biological behavior is supported by the data of a low Ki-67 labeling index and a lack of p53 or c-erbB-2 protein over-expression. Because of its resemblance to normal gastric mucosa or mucosa with intestinal metaplasia, EWDA is often misdiagnosed. To prevent the misdiagnosis of such lesions, the clinical and pathologic characteristics should be taken into consideration. 展开更多
关键词 Stomach neoplasms Extremely well-differen-tiated adenocarcinoma Ki-67 p53 c-erbB-2
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Immunohistochemical study on distribution of endocrine cells in gastrointestinal tract of flower fish (Pseudophoxinus antalyae) 被引量:6
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作者 Kenan lnar Nurgül ■enol M Rü■tü zen 《World Journal of Gastroenterology》 SCIE CAS CSCD 2006年第42期6874-6878,共5页
AIM: To detect distribution and relative frequency of endocrine cells in gastrointestinal tract of flower fish (Pseudophoxinus antalyae). METHODS: The intestinal tract of flower fish was divided into four portions... AIM: To detect distribution and relative frequency of endocrine cells in gastrointestinal tract of flower fish (Pseudophoxinus antalyae). METHODS: The intestinal tract of flower fish was divided into four portions from proximal to distal; the enlarged area after oesophagus and anterior, middle and posterior intestine. Immunohistochemical method using the peroxidase anti-peroxidase complex was employed. All antisera between four portions of flower fish were compared using ANOVA. RESULTS: Eleven types of gut endocrine cells were determined; they were immunoreactive for calcitonin gene related peptide, substance P, vasoactive intestinal peptide, bombesin, somatostatin-14, secretin, TrkA, TrkB, TrkC, neurotensin, neuropeptide Y, which were found in almost all portions of the gastrointestinal tract. CONCLUSION: The regional frequency of immunoreactive distribution and relative cells in the flower fish, Pseudophoxinus antalyae, are essentially similar to those of other fish. 展开更多
关键词 Flower fish Gastrointestinal tract Immunoreactive cells Pseudophoxinus antalyae
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Tropomyosin is localized in the nuclear matrix and chromosome scaffold of physarum polycephalum 被引量:1
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作者 ZENG XIAN LU MING DA JIAO +2 位作者 MIAO XING XIAO GUANG WANG SHUI HAO(Institute of Genetics and Cytology, Northeast Normal University, Changchun 130024, China)(Department of Biology, Changchun Normal College,Changchun 130032, China) 《Cell Research》 SCIE CAS CSCD 1999年第1期61-69,共9页
The nuclei and chromosomes were isolated from plasmodia of Physarum polycephalum. The nuclear matrir and chromosome scaffold were obtained after the DNA and most of the proteins were extracted with DNase I and 2 M NaC... The nuclei and chromosomes were isolated from plasmodia of Physarum polycephalum. The nuclear matrir and chromosome scaffold were obtained after the DNA and most of the proteins were extracted with DNase I and 2 M NaCl. SDS-PAGE analyses revealed that the nuclear matrir and chromosome scaffold contained a 37 kD polypeptide which is equivalent to tropomyosin in molecular weight. Immunofluorescence observations upon slide preparations labeled with anti-tropomyosin antibody showed that the nuclear matrix and chromosome scaffold emanated bright fluorescence, suggesting the presence of the antigen in them.Immunodotting results confirmed the presence of tropomyosin in the nuclear matrix and chromosome scaffold. Immunoelectron microscopic obserwtions further demonstrated that tropomyosin was dispersively distributed in the interphase nuclei and metaphase chromosomes. 展开更多
关键词 Chromosome scaffold nuclear matrix Physarum polycephalum TROPOMYOSIN
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The mitosis and immunocytochemistry of olfactory ensheathing cells from nasal olfactory mucosa 被引量:8
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作者 刘锦波 唐天驷 +2 位作者 龚爱华 盛伟华 杨吉成 《Chinese Journal of Traumatology》 CAS 2005年第5期306-310,共5页
Objective: To culture olfactory ensheathing cells (OECs) of rats in vitro and to investigate its morphology, mitosis and immunocytochemistry, and to explore if the OECs could be a new donation for transplantation. ... Objective: To culture olfactory ensheathing cells (OECs) of rats in vitro and to investigate its morphology, mitosis and immunocytochemistry, and to explore if the OECs could be a new donation for transplantation. Methods: OECs were harvested from olfactory mucosa of Sprague Dawleys rats based on the differing rates of attachment of the various cell types, followed by glial fibrillary acidic protein (GFAP), nerve growth factor ( NGF), anti-low affinity receptor for NGF ( NGFRp75 ), brain-derived neurotrophic factor (BDNF) , neurotrophino3 ( NTo3 ) and S-100 immunocytochemistry. The morphological changes and mitosis were observed under a phase contrast microscope at different culture time. Results - Three morphologically distinct types of cells, bipolar, multipolar and flat morphology were present in the primary culture of adult rat olfactory mucosa. Mitosis was characterized by a retraction of all processes, forming a sphere that divided into spherical daughter cells, the daughter cells sent out their processes. The OECs were immunoreactive for GFAP, NGFRp75, S-100, NGF, BDNF and NT-3. Conclusions: The OECs from nasal olfactory mucosa cultivated in the medium with fetal bovine serum could survive, divide, differentiate, and express the neurotrophin. It may become an accessible source for autologous grafting in spinal cord injury. 展开更多
关键词 Olfactory mucosa Immunohistochemistry: Olfactory ensheathing cells
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Essential sequence of the N-terminal cytoplasmic localization-related domain of huntingtin and its effect on huntingtin aggregates 被引量:4
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作者 YAN YaPing PENG DanTao +6 位作者 TIAN Jun CHI JingWei TAN JieQiong YIN XinZhen PU JiaLi XIA Kun ZHANG BaoRong 《Science China(Life Sciences)》 SCIE CAS 2011年第4期342-350,共9页
Huntington's disease (HD) is caused by abnormal CAG repeat expansion in the 5'-end of the Huntingtin (HTT) gene. In addition to the canonical C-terminal full-length huntingtin (htt) nuclear export signal, a cy... Huntington's disease (HD) is caused by abnormal CAG repeat expansion in the 5'-end of the Huntingtin (HTT) gene. In addition to the canonical C-terminal full-length huntingtin (htt) nuclear export signal, a cytoplasmic localization-related domain (CLRD) in the N-terminus of htt has recently been reported. Here, we analyzed this domain by introducing deletion and substitution mutations in a truncated N-terminal htt protein and subsequently monitored htt expression, aggregation and subcellular localization by immunocytochemistry and Western blot analysis. We demonstrated that Htt1-17 was the essential sequence for htt cytoplasmic localization. We also found that the subcellular distribution of htt was altered when Htt1_17 was mutated to contain amino acids of different charges, suggesting a structural requirement of Htt1-17 for the cytoplasmic localization of htt. Deletion of the first three amino acids did not affect its association with mitochondria. We observed that defective cytoplasmic localization resulted in a reduction of total htt aggregates and increased nuclear aggregates, indicating that the subcellular distribution of the protein might influence the aggregation process. These studies provide new insight into the molecular mechanism of htt aggregation in HD. 展开更多
关键词 HUNTINGTIN AGGREGATES cytoplasmic localization related domain mitochondria POLYGLUTAMINE
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