中国水产科学2018，Vol.25Issue(2)：346-353,8.DOI:10.3724/SP.J.1118.2018.17211
栉孔扇贝心脏细胞的体外培养
Cell cultures from Chlamys farreri heart cells
摘要
Abstract
Cells cultured in vitro play an important role in organism-resource protection and functional identifica-tion of genes, as well as mechanism exploration and prevention of animal disease. However, tissue cells that can survive for a long time in vitro are limited at present in marine shellfish. In this study, primary cultures of heart cells from the scallop Chlamys farreri were started using an explant method,and a primary culture system-which can keep the cells alive for a long time in vitro-was established using an optimizing-medium method.Effects of three basic media (L-15, M199, and L-15+M199) on the number of migrated cells from the explants and cell sur-vival time were compared;the L-15 medium was verified to be the optimal basic medium for C.farreri heart cells. Furthermore, the optimal supplemental-factor combination for C.farreri heart cells was the L-15 medium sup-plemented with 5% FBS, 50 mmol/L taurine and 6 mmol/L Ca2+(orthogonal experiment of three factors and three levels),in which cardiomyocytes could survive for 2 months in vitro.In this study,the results show that low con-centration serum (5% FBS) was most favorable for the migration of cells from the tissue mass. With an increase in FBS concentration, the cell-migration ability was weaker, and the survival time of primary cells became shorter. After the addition of taurine, the survival time of primary cells was significantly prolonged. However, the Ca2+(2 mmol/L,4 mmol/L,and 6 mmol/L)had no significant effect on C.farreri heart-cell cultures.Perhaps the additive taurine regulated intracellular Ca2+to a suitable level, resulting in the addition of Ca2+without significant im-provement in the primary culture. In this primary culture, most cells were cardiomyocytes; part of the cardiomyo-cytes beat at regular intervals within a short time, and cardiomyocytes and myotubes were formed in part of the region.This phenomenon was also reported in the primary culture of heart cells of Crassostrea gigas.This study provides a useful foundation for further studies on C.farreri basic biology and functional genes.关键词
栉孔扇贝/心脏细胞/体外培养/正交实验Key words
Chlamys farreri/heart cells/cell culture in vitro/orthogonal design分类
生物科学引用本文复制引用
季爱昌,王华,晏萌,张志峰..栉孔扇贝心脏细胞的体外培养[J].中国水产科学,2018,25(2):346-353,8.基金项目
国家863计划项目(2012AA10A402). （2012AA10A402）
