CTLA4.FasL免疫抑制效应及对肝前体细胞增殖分化潜能的影响

目的探讨融合蛋白CTLA4.FasL对肝前体细胞(LEPCs)增殖分化潜能的影响及抑制异种排斥反应的效应。方法克隆CTLA4.FasL基因,构建携带CTLA4.FasL基因与红色荧光蛋白(mCherry)双顺反子结构的重组慢病毒载体Lv-CTLA4.FasL-IRES-mCherry。优化慢病毒感染LEPCs的条件,建立高表达CTLA4.FasL的LEPCs,荧光显微镜观察mCherry的表达,Western blot检测CTLA4.FasL的表达,酶联免疫吸附测定法(ELISA)测定细胞培养上清中CTLA4.FasL的浓度,水溶性四氮唑(WST-1)法检测细胞的增殖活性,Real time PCR(RT-PCR)检测干细胞相关基因CK19和c-Kit mRNA的表达,5-溴脱氧尿嘧啶核苷(Brdu)掺入法测定CTLA4.FasL-LEPCs在异种混合淋巴细胞培养体系中对大鼠淋巴细胞增殖的抑制作用。结果构建重组慢病毒载体Lv-CTLA4.FasL-IRES-mCherry,病毒滴度为2×108 TU/mL。在感染复数(multiplicity of infection,MOI)为10,聚凝胺质量浓度是5μg/mL时,感染效率约90%。Western blot证实了CTLA4.FasL的表达,在细胞培养上清中其质量浓度约为(0.72±0.10)μg/mL。CTLA4.FasL-LEPCs细胞增殖活性未受到影响,CK19和c-Kit基因mRNA表达水平无变化。CTLA4.FasL-LEPCs细胞可显著抑制大鼠淋巴细胞的增殖(P<0.05)。结论构建成功的重组慢病毒载体Lv-CTLA4.FasL-IRES-mCherry可介导CTLA4.FasL基因在LEPCs中高效表达;CTLA4.FasL可有效地抑制异种排斥反应,同时不损害LEPCs增殖活性和分化潜能。     

HAIR CELL-LIKE CELL GENERATION INDUCED BY NATURE CULTURE OF ADULT RAT AUDITORY EPITHELIUM

Objective To establish adult rat auditory epithelial cell culture and try to find precursor cells of auditory hair cells in vitro. Methods With refinement of culture media and techniques, cochlear sensory epithelial cells of adult rat were cultured. Immunocytochemistry and Bromodeoxyuridine （BrdU） labeling were used to detect properties and mitotic status of cultured cells. Results The cultured auditory epithelial cells showed a large, flat epithelial morphotype and expressed F-actin and cytokeratin, a subset of cells generated from auditory epithelium were labeled by calretinin, a specific marker of early hair cell. Conclusion Adult rat auditory epithelium can be induced to generate hair cell-like cells by nature culture, this phenomenon suggests that progenitor cells may exist in rat cochlea and they may give birth to new hair cells. Whether these progenitor cells are tissue specific stem cells is still need more study.     

XY 轰轰烈HL150-10F

也许大家看到本文的题目会心生疑问,这个"XY"是试车文章的题目么?下面就让笔者给您具体分析这"XY"与轰轰烈HL150-10F的缘由吧!XY是一个基因组合。人体细胞中有23对染色体即DNA的聚合体,男性的染色体组成是X和Y,女性的是X和X。也就是说,XY是所有男性共有的,与生俱来的特质。就像是摩托车和女人一样,是所有男人都有的梦想。     

铁皮石斛早期体细胞胚胎发育及原球茎形态发生研究

铁皮石斛（Dendrobium candidum Wall ex Lindl）是兰科石斛属多年生草本植物,是一种濒危物种,我们以此为材料建立了高效体细胞胚胎（以下简称“体胚”）发生体系,并从含体胚的胚性愈伤组织中分离出一些胚胎发生的特异基因。本项目就是在此基础上,欲建立早期体胚分离纯化技术,     

体细胞核转移

经过两年多透彻的伦理性及科学性检查之后．身处哈佛与波士顿儿童医院的哈佛千细胞研究所（HSCI）研究员已经获许开展使用体细胞核转移（SCNT）的实验来创建疾病特异干细胞系。其目标是开发出新的疗法，使大量患有目前不能治愈疾病的人们免遭痛苦。[第一段]