| 洪宇航,杨筱珍,张金彪,梁攀,赵柳兰,成永旭.2012.中华绒螯蟹血细胞原代培养条件的优化.动物学杂志,47(2):52-58. |
| 中华绒螯蟹血细胞原代培养条件的优化 |
| The Optimization of Primary Hemocyte Culture in the Mitten Crab,Eriocheir sinensis |
| 投稿时间：2011-10-24 修订日期：2012-01-04 |
| DOI： |
| 中文关键词: 中华绒螯蟹 血细胞 原代培养 |
| 英文关键词:Eriocher sinensis Hemocyte Primary culture |
| 基金项目:国家自然科学基金项目(No.30700609,30871927),上海市中华绒螯蟹产业技术体系项目(No.D2010100208),上海高校创新团队(第二期)项目(水产动物营养饲料与养殖环境); |
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| 中文摘要: |
| 比较了几种常见血细胞培养基(L-15、2×L-15、3×L-15、M199和RMPI-1640)对中华绒螯蟹(Eriocheir sinensis)血细胞原代培养中细胞形态以及存活率的影响,以及在筛选获得的最佳培养基中添加不同比例胎牛血清(FBS)(0%、5%、10%和15%),进一步观察了血清对中华绒螯蟹血细胞培养效果的比较。结果表明,3×L-15培养基培养效果较好,所培养的细胞形态相对完整,数量较多,培养至96 h时血细胞存活率仍大于60%;而其他4种培养基效果较差,培养12 h存活率均低于50%,且细胞形态结构变化明显。以3×L-15培养基为基础,添加不同比例胎牛血清后发现,对细胞存活有显著影响,存活率明显降低。因此,不添加血清的3×L-15培养基对中华绒螯蟹血细胞的生长较为适宜。 |
| 英文摘要: |
| In the present study,the effects of several frequently-used cell culture media (L-15,2×L-15,3×L-15,M199 and RMPI-1640) on cell morphology and viability of hemocytes from the mitten crab,Eriocheir sinensis were studied,in order to screen an optimum culture medium for crab hemocytes.Furthermore,different proportions of fetal bovine serum (FBS) (0%,5%,10%,and 15%) as a supplement were added into an optimum culture medium to research the effects of serum on hemocyte culture.The results showed that:3×L-15 culture medium could support the best survival of hemocytes in in vitro culture.The hemocytes remained a maximum number and intact morphology relatively,and the viability of hemocytes after 96 h culture was more than 60% in 3×L-15 medium.On the contrary,the other media did not support well of the hemocyte culture,with the viability decrease to lower than 50% and obvious cell morphology changes within 12 h.In addition,the supplement of FBS had adverse effect on cell morphology and viability.These results show that 3×L-15 medium without FBS is adequate for E.sinensis primary hemocyte culture. |
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