水产学报2018，Vol.42Issue(4)：513-521,9.DOI:10.11964/jfc.20170310756
团头鲂骨组织细胞系的建立及其生物学特性
Establishment and characterization of bone cell line for blunt snout bream(Megalobrama amblycephala)
摘要
Abstract
In order to study the characteristics of fish bone cells, methods for isolating and culturing bone cells from caudal vertebrae and rib of blunt snout bream Megalobrama amblycephala were explored in the present study, and the biological characters of bone cells were identified. The bone cells were cultured with tissue culture and trypsin digestion respectively by using 3-month-old M.amblycephala caudal vertebrae and ribs from tail part. The optimal conditions were as follows: 25 ng/mL epidermal growth factor (EGF) and 25 ng/mL basic fibroblast growth factor (bFGF) were added to the M-199 medium at 28 °C using tissue culture. The results of cell biology identification showed that the cultured cells had the biological activity of typical osteoblasts, with alkaline phos-phatase staining, calcium nodules alizarin red staining and mineralized nodules von Kossa staining being positive. The osteocalcin content was 997.25 ng/L in the bone cells tested by the fish bone osteocalcin ELISA kit. The genes of runx2a,runx2b and osterix which regulated osteoblast differentiation,were expressed in bone cells,with the runx2b and osterix expression levels significantly higher than that in muscle cells.Results indicated that bone cells of fish have good proliferation and faster growth rate as well as stable cell biological characteristics, which is suit-able for related studies in vitro.关键词
团头鲂/骨组织细胞/细胞系/基因表达/生物学特性Key words
Megalobrama amblycephala/bone cells/cell line/gene expression/biological characterization分类
生物科学引用本文复制引用
关柠楠,兰天,陈宇龙,刘莉芳,周芬,高泽霞..团头鲂骨组织细胞系的建立及其生物学特性[J].水产学报,2018,42(4):513-521,9.基金项目
国家自然科学基金(31472271) （31472271）
武汉市青年科技晨光计划(2016070204010143) （2016070204010143）
国家大宗淡水鱼类产业技术体系建设项目(CARS-45-08) （CARS-45-08）
国家级大学生创新创业训练计划(2015308200630)National Natural Science Foundation of China (31472271) （2015308200630）
Wuhan Youth Science and Techno-logy Plan (2016070204010143) （2016070204010143）
Modern Agriculture Industry Technology System Construction Projects of China Titled as-Staple Freshwater Fishes Industry Technology System (CARS-45-08) （CARS-45-08）
National Undergraduate Innova-tion and Entrepreneurship Training Program (2015308200630) （2015308200630）
