中国水产科学Issue(2)：225-234,10.DOI:10.3724/SP.J.1118.2014.00225
三角帆蚌外套膜细胞体外培养优化及体内植入培养对细胞生长的影响
Impact of in vitro optimization and in vivo implantation culture on the growth of Hyriopsis cumingii mantle cells
摘要
Abstract
In China, Hyriopsis cumingii is the shellfish of choice for cultivating freshwater pearls;however, there has been very little research on cell and mantle tissue culture. Based on Dulbecco’s modified Eagle’s medium (DMEM), this study attempted to optimize modified formula of medium and buffer for mantle tissue and cell culture. The ratio of RNA/DNA as an evaluation index of cell proliferation was determined and, using microscopic observations, tissue cul-ture cells versus time, speed and in vitro cell energy were established. Hoechst fluorescent staining of DNA was carried out using 24-, 108-and 120-h cells of in vitro culture, and then the three marked cell groups were implanted into the mantle for culture in vivo. The results show that, after optimization, the buffer and medium improved cell migration from the mantle, the velocity of migration and the significantly increased cell number (P<0.05).Cell vitality also in-creased with incubation time. Cell vitality was maximal for culture in vitro at 108 h, with a RNA/DNA ratio of 24.53, but was followed by a significant decline (P<0.05).Microscopic observations of cells were in agreement with the deter-mined RNA/DNA index values. When in vitro cultured cells were implanted into the mantle a second time, in vitro ac-tivity improved and culture conditions increased cell activity, with RNA/DNA values up to 25.45, but vitality declined significantly after 168 h when injected into the cell for in vivo culture (P<0.05). Regarding the poor in vitro activity of cells, it would appear that a living environment has no significant effect on cell viability. Our study provides basic but valuable information for further research on mantle cell proliferation and the establishment of cell lines.关键词
三角帆蚌/外套膜/细胞培养/组织培养/RNA/DNA活力检测/淡水珍珠Key words
Hyriopsis cumingii/cell culture/mantle/tissue culture/RNA/DNA/fresh water pearl分类
农业科技引用本文复制引用
李倩,施志仪,李文娟,黄凯,祁晓翔..三角帆蚌外套膜细胞体外培养优化及体内植入培养对细胞生长的影响[J].中国水产科学,2014,(2):225-234,10.基金项目
国家自然科学基金资助项目(31201991) （31201991）
上海市博士后科研资助计划面上项目(09R21413200) （09R21413200）
教育部博士点基金项目(20123104120003) （20123104120003）
上海市优秀青年项目(ssc11004) （ssc11004）
上海市重点学科水生生物学建设项目(S30701) （S30701）
上海市高校知识服务平台项目(ZF1206) （ZF1206）
上海海洋大学博士启动基金资助项目 （）
