PDF HTML阅读 XML下载 导出引用 引用提醒 栉孔扇贝心脏细胞的体外培养 DOI: 作者: 作者单位: 中国海洋大学 海洋生物遗传育种教育部重点实验室, 山东 青岛 266003 作者简介: 季爱昌(1987-),女,博士研究生,主要从事海洋动物细胞生物学研究.E-mail:jiaich@126.com 通讯作者: 中图分类号: Q751;S968 基金项目: 国家863计划项目(2012AA10A402). Cell cultures from Chlamys farreri heart cells Author: Affiliation: Key Laboratory of Marine Genetics and Breeding, Ministry of Education;Ocean University of China, Qingdao 266003, China Fund Project: 摘要 | 图/表 | 访问统计 | 参考文献 | 相似文献 | 引证文献 | 资源附件 | 文章评论 摘要:体外培养细胞对研究生物的资源保护、功能基因及病害发生机理与防治等均具有重要意义。然而,目前海洋双壳贝类中可以长期存活的组织细胞是有限的。本研究采用植块法启动栉孔扇贝()心脏细胞的原代培养,通过优化培养基的方法,建立了可使其长期存活的原代培养体系。根据组织块迁出细胞的数量和细胞存活时间,确定L-15是3个培养基(L-15,M199,L-15+M199)中最适宜栉孔扇贝心脏细胞的基础培养基。通过三因子三水平正交实验,得出栉孔扇贝心脏细胞适宜的添加物组合:L-15基础培养基中添加5%胎牛血清、50 mmol/L牛磺酸和6 mmol/L Ca2+。原代培养产物中以心肌细胞为主要的细胞类型,其中部分心肌细胞在原代培养14 d内可进行节奏性搏动;部分细胞可局部形成心肌束和肌管样结构;心肌细胞在体外存活时间达2个月。本研究将为栉孔扇贝基础生物学和功能基因的研究提供细胞平台。 Abstract:Cells cultured play an important role in organism-resource protection and functional identification of genes, as well as mechanism exploration and prevention of animal disease. However, tissue cells that can survive for a long time are limited at present in marine shellfish. In this study, primary cultures of heart cells from the scallop were started using an explant method, and a primary culture system-which can keep the cells alive for a long time -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 survival time were compared; the L-15 medium was verified to be the optimal basic medium for heart cells. Furthermore, the optimal supplemental-factor combination for heart cells was the L-15 medium supplemented 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 this study, the results show that low concentration 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 heart-cell cultures. Perhaps the additive taurine regulated intracellular Ca2+ to a suitable level, resulting in the addition of Ca2+ without significant improvement in the primary culture. In this primary culture, most cells were cardiomyocytes; part of the cardiomyocytes 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 This study provides a useful foundation for further studies on basic biology and functional genes. 参考文献 相似文献 引证文献