文昌鱼鳃组织原代培养  

Primary cell culture from gill explants of amphioxus

作　　者：石松林[1] 黄晓明[1] 李祺福[1] 王三英[1] 杨海波[1] 

机构地区：[1]厦门大学生命科学学院,福建厦门361005

出　　处：《海洋科学》2009年第3期44-48,共5页Marine Sciences

基　　金：国家863计划资助项目(2005AA626013)

摘　　要：采用组织块培养法培养文昌鱼（Branchiostoma belcheri japonicum）鳃细胞，使用L15，M199，Eargle’s MEM为基础培养基，加入一定量的NaCl及5％，10％，20％等不同体积分数的牛血清。经条件优化后得到最适合的培养条件为2L15＋20％FBS＋0．45％NaCl，在26～28℃条件下培养，组织决贴壁后12h内就有至少4类上皮样细胞从中迁移出来，这些细胞在前3天迅速增殖，到第6天基本上达到平稳期，往后细胞开始凋亡，最长可以存活近两个星期。鳃细胞的原代培养为文昌鱼细胞体外培养的深入研究提供了实验依据。Primary cultures of gill cells were initiated from gill filament explants of amphioxus （Branchiostoma belcheri japonicum） by a tissue block culture method. The explants were cultured in Leibovitz L15 medium, M199, Eargle＇s MEM medium with 5 %, 10% or 20 % foetal calf serum（FCS） and a certain amount of NaCl. We found that gill cell could grow better in 2 L15 medium than in others. The attachment efficiency was serum-dependent though increased FCS concentration did not stimulate further outgrowth of cells. The explants produced four different kinds of epithelia cell outgrowth in 12 h after attachment as a sheet of cells. There was high proliferation for the first 3 days, then a stable plateau for 3 days followed by a decline phase for 14 days. The use of gill explants to establish primary cultures of fish gill cells has advantages which include longevity of the culture and successive proliferations from explants and could provide a useful tool for the investigation of long-term processes in cellular biology.

关 键 词：文昌鱼(Branchiostoma BELCHERI japonicum) 鳃组织 原代培养 

分 类 号：Q813.11[生物学—生物工程] X503.225[环境科学与工程—环境工程]