安徽农业科学2016，Vol.44Issue(15)：130-134,5.
斑马鱼性腺组织的原代细胞培养技术的建立
Development of Primary Cell Cu lture from the Gonad Tissues of Zebrafish
摘要
Abstract
Objective] To develop a primary cell culture system for the cells from ovary and testis tissues of zebrafish and to obtain the corre -sponding cell monolayer of both primary culture and subculture.[Method]Primary cell cultures were derived using the explants culture method. The primary cells were sub-cultured by 0.25% trypsin digestion.[Result] The zebrafish gonad-derived cells were cultured in DMEM/F12 (pH 7.0-7.2) at an optimal temperature of 24 ℃.The cells had better growth and survivor when the medium mentioned above was supplemented with 20 ng/mL epidermal growth factor (EGF),20 ng/mL basic fibroblast factor (bFGF),0.5 mmol/Lβ-mercaptoethanol and 15%fetal bovine serum (FBS).For ovary tissues,epithelium-like cells first migrated from the explants at 3~4 days after seeding.These epithelium-like cells could only survive for about 5 days in vitro,and then tended to apoptosis.However, active migration of fibroblast-like cells and another kind of epitheli-um-like cells were then observed and a primary cell monolayer with 80%confluence could be formed within10 days.Then the primary cell mono-layer could be successfully sub-cultured once.The epithelium-like cells can’t re-attach and die soon after subculture.However,the re-attached fi-broblast-like cells could healthily survive for another 7 days and then tend to apoptosis.No obvious cell division was observed in both the primary and subculture cells.For testis tissues,both epithelium-like cells and fibroblast-like cells migrated from the explants at 5 days after seeding.At 16 days after seeding,a primary cell monolayer with 60%confluence was obtained and then tended to apoptosis.[Conclusion] This study has estab-lished a primary cell culture system suitable for the gonad tissues of zebrafish ,and the fibroblast-like cell monolayer derived from the ovary tissues has been successfully sub-cultured for one time.关键词
斑马鱼/性腺组织/原代细胞培养/传代培养Key words
Zebrafish/Gonad tissue/Primary cell culture/Subculture分类
农业科技引用本文复制引用
董丹丹,孙燕侠,郭华荣..斑马鱼性腺组织的原代细胞培养技术的建立[J].安徽农业科学,2016,44(15):130-134,5.基金项目
国家自然科学基金项目（31472274，31172391）；海洋经济创新发展区域示范项目（12 PYY001 SF08）；中国海洋大学本科生SRDP项目。 （）
