克氏原螯虾精巢与输精管组织细胞原代培养方法初探  

Preliminary Studies on the Cell Culture of the Spermary and Spermaduct in Procambarus clarkii

作　　者：刘凯于[1] 姚汉超[1] 邱宝国[1] 郭建军[1] 彭建新[1] 洪华珠[1] 

机构地区：[1]华中师范大学生命科学学院教育部农药与化学生物学重点实验室,武汉430079

出　　处：《动物学杂志》2007年第4期60-64,共5页Chinese Journal of Zoology

基　　金：湖北省自然科学基金项目(No9523);华中师范大学生命科学学院自然科学基金项目

摘　　要：将剪碎的克氏原螯虾(Procambarus clarkii)精巢与输精管组织块分别接种于含20%FBS、双抗以及添加了适量无机盐等的M199-MK、MEM、S20、Grace’s和L-15培养基中,25℃恒温静置培养。结果表明,在Grace’s培养基中,只有少量组织块贴壁。在其余培养基中,大多数组织块能在1～2周内逐渐贴壁,L-15和S20培养基培养效果最好。组织块贴壁后,一些成纤维细胞和上皮样细胞逐渐从组织块迁移出来。传代后组织块可继续贴壁并重新迁移出一些成纤维细胞与少量上皮样细胞。添加部分草鱼细胞系PSF或斜纹夜蛾细胞系Sl驯化的培养基进行培养,驯化培养基:新鲜培养基=1:5(v/v),结果表明,Sl驯化培养基能促进细胞的迁移。组织块一般可离体培养2～4个月,但是都未能形成细胞单层。胰蛋白酶消化后分散的虾细胞不能贴壁。The minced tissue of the spermary and spermaduct in Procambarus clarkii was cultured in different media including M 199- MK, MEM, S20, Grace＇ s medium and modified leibovitz L-15 medium, supplemented with 20 % FBS, antibiotics,and salts at 25℃ .The results showed that only a few pieces of tissue attached to the flasks in Grace＇s medium;many pieces of tissue could attach to the culture flasks during 1 to 2 weeks in M199-MK and MEM media; and the cells grew well in S20 medium or modified L-15 medium .Two types of conditioned media were tested and the modified L-15 medium conditioned by insect cell line S1 could improve the migration of the cells. The cells could survive for 2 - 4 months, but could not form a monolayer. The cells dissociated by trypsin could not attach to the flasks.

关 键 词：克氏原螯虾 细胞培养 精巢 输精管 原代培养 

分 类 号：S917.4[农业科学—水产科学]