中华绒螯蟹血细胞原代培养条件的优化

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。

The Optimization of Primary Hemocyte Culture in the Mitten Crab,Eriocheir sinensis

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.