仿刺参体腔细胞原代培养方法初探

The primary culture methods of coelomocytes in sea cucumber Apostichopus japonicus

以仿刺参Apostichopus japonicus体腔细胞为材料,分别用体腔液和MEM、L-15两种培养基进行原代培养,初步探讨了仿刺参体腔细胞培养的基本条件。结果表明:在18℃下封闭培养,体腔细胞在体腔液中直接培养能够贴壁、伸展,细胞呈球形和上皮样,但凝集现象严重;在合成培养基(MEM、L-15)中,体腔细胞凝集明显减少;与MEM培养基相比,体腔细胞在L-15培养基中贴壁时间长,伸展状态好,上皮样细胞较多,可培养至72 h;培养基中血清的添加量与细胞的培养状态和可培养时间关系不明显。

The coelomocytes in sea cucumber Apostichopus japonicus were primarily cultured in the sea cucumber coelomic fluid, MEM and L - 15 medium to evaluate the coelomocyte culture conditions. The results showed that the coelomocytes directly cultured in the coelomic fluid showed attachment and extension, with a strongly agglutination under closed culture at 18 ℃, and the cells were shaped like a sphere or skin. The coelomocytes cultured in the compound medium （ MEM, L - 15 ） showed less agglutination. The coelomocytes in L - 15 medium had a relatively long period of attachment and a better stretching fettle, and were cultured in 72 hours compared with MEM medium. There were no significant correlation between the culturing period and cell vitality and the serum concentration.