PDF(Pubmed)
Abstract：
Cell culture provides useful model systems used in a wide range of biological applications, but its utility in marine invertebrates is limited due to the lack of immortalised cell lines. Primary cell and tissue cultures are typically used but remain poorly characterised for oysters, which can cause issues with experimental consistency and reproducibility. Improvements to methods of repeatable isolation, culture, and characterisation of oyster cells and tissues are required to help address these issues. In the current study, systematic improvements have been developed to facilitate the culture of primary cells from adult Pacific oyster tissues and identify novel cell morphologies that have not been reported previously. Cultures analysed by light microscopy, qPCR, and live cell imaging demonstrated maintenance of live, metabolically active Pacific oyster cells for several weeks post-explant. Interestingly, whole hearts dissected from adult oysters were found to continue contracting rhythmically up to 8 weeks after being transferred to a tissue culture system. Mantle tissue explants were also actively moving in the culture system. These improvements in primary cell culture of bivalves may be beneficial for research in ecotoxicology, virology, immunology, and genetic resistance to disease.
摘要：
细胞培养提供了在广泛的生物应用中使用的有用的模型系统，但是由于缺乏永生化的细胞系，其在海洋无脊椎动物中的应用受到限制。通常使用原代细胞和组织培养物，但对牡蛎的特征仍然很差，这可能会导致实验一致性和可重复性的问题。可重复隔离方法的改进，文化,牡蛎细胞和组织的特征需要帮助解决这些问题。在目前的研究中,已经开发了系统的改进，以促进从成年太平洋牡蛎组织中培养原代细胞，并确定以前没有报道的新细胞形态。通过光学显微镜分析培养物，qPCR,活细胞成像显示了活的维持，外植体后几周代谢活跃的太平洋牡蛎细胞。有趣的是,从成年牡蛎解剖的整个心脏在转移到组织培养系统后持续有节奏地收缩长达8周。地幔组织外植体也在培养系统中积极移动。双壳类原代细胞培养的这些改进可能有利于生态毒理学研究。病毒学,免疫学,和对疾病的遗传抗性。
