Establishment and characterization of a new cell line from the kidney of spotted halibut <Emphasis Type="Italic">Verasper variegates</Emphasis>

A cell line,SHK,was derived from the kidney of spotted halibut Verasper variegates.The cell line was subcultured more than 40 passages in minimum essential medium(MEM)supplemented with fetal bovine serum(FBS)and 10 ng ml-1 basic fibroblast growth factor(bFGF).Cell morphology from primary culture and subculture was observed continuously by microscopy.The SHK cell line consisted predominantly of fibroblast-like cells.The cell line was able to grow between 20℃ and 30℃ with the optimum growth at 24℃ and with a reduced growth between 12℃ and 20℃.The growth rate of the cells increased as the proportion of FBS increased from 10% to 20% at 28℃ with optimum growth at the concentration of 20%.The doubling time of the cells was determined to be 44.8 h.Chromosome analysis revealed that 52% of the SHK cells maintained a normal diploid chromosome number (2n=46).The cells were successfully transfected with green fluorescent protein(GFP)reporter plasmids and the expression of GFP gene in the cells indicated the possible utility of the cells in gene expression studies.The cells were infected by lymphosystis disease virus(LCDV)and found to be susceptible to the virus in cytopathic effect(CPE)observation.The infection was confirmed by PCR and electron microscopy experiments,which proved the existence of the viral particles in the cytoplasm of the virus-infected cells.     

沙蜇和白色霞水母刺丝囊毒素的细胞毒性比较研究

分别自沙蜇和白色霞水母提取刺丝囊毒素蛋白,以小鼠皮下结缔组织细胞(A9)和人横纹肌瘤细胞(A-673)为受试对象,比较两种水母的刺丝囊毒素蛋白对其的细胞毒性,以期为评估和管理两种水母的毒害风险提供科学依据。结果显示:沙蜇和白色霞水母刺丝囊毒素对培养中的A9和A-673细胞存活率都具有抑制作用,且该抑制作用呈显著的时间和剂量依赖性。白色霞水母刺丝囊毒素对A9和A-673细胞毒性的半数效应浓度(如:对A9和A-673的EC50-24h分别为49.8μg/mL和43.6μg/mL)低于沙蜇刺丝囊毒素的半数效应浓度(如:对A9和A-673的EC50-24h分别为78.0μg/mL和70.1μg/mL);两种水母毒素对A-673或A9的细胞毒性强于已报道的根口类和旗口类水母毒素对哺乳动物肺、肝、血管平滑肌、神经、乳腺等的细胞毒性。由此可见,白色霞水母刺丝囊毒素的细胞毒性更强,横纹肌、结缔组织对水母刺丝囊毒素作用敏感。研究结果对于评估水母毒害风险和制定防护对策有重要意义。     

海洋无脊椎动物细胞培养与细胞干性的捕获

细胞系的构建在生物学领域具有重要意义,与其他动物(如昆虫)不同,经过数十年的发展,海洋无脊椎动物细胞研究仍停留在原代培养水平,没有建立起克隆化的永生细胞系.本文主要介绍了近年来海洋无脊椎动物细胞培养的研究进展,还介绍了一个基本原理,即海洋无脊椎动物永生细胞系的建立,可以通过捕获细胞的干性而实现.捕获细胞干性的第一条途径...     

异育银鲫(Carassius auratus gibelio)脊髓组织细胞系的建立及对CyHV-2的敏感性

鲫造血器官坏死病是近些年来危害养殖鲫鱼最为严重的传染性疾病,其病原为鲤疱疹病毒Ⅱ型(Cyprinid herpesvirus 2, CyHV-2)。本研究以异育银鲫(Carassius auratus gibelio)脊髓组织为材料,进行组织细胞体外培养,构建对鲤疱疹病毒Ⅱ型敏感的异育银鲫脊髓细胞系(Spinalcordtissuecell lines of Carassius auratus gibelio, CSC)。采用组织块移植法,进行异育银鲫脊髓细胞的原代培养,并进行传代,已传代至第128代次,获得稳定传代的细胞系后,采用低渗法进行细胞染色体核型分析,并进行鲤疱疹病毒Ⅱ型的敏感性测试与传代。结果表明,以含10%胎牛血清的L-15培养基,在24℃恒温条件下,可稳定传代培养异育银鲫脊髓细胞,传代细胞呈成纤维样或纺锤状;核型分析显示,染色体数为156±2条,该细胞为三倍体细胞。通过对CyHV-2敏感性的测试,CSC细胞感染CyHV-2后产生典型的细胞病变,感染第7天的病变细胞液的TCID₅₀达到10^9.33/mL,CyHV-2病毒可在CS...