Developments in marine invertebrate primary culture reveal novel cell morphologies in the model bivalve Crassostrea gigas

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.

Cell culture and genetic transfection methods for the Japanese scallop, Patinopecten yessoensis

“…The morphology of cultured scallop cells varied among organs. In both types of gonadal cells, most cultured cells were fibroblast-like, consistent with previous findings [9]. In this study, round adhered cells and spread adhered cells were also observed in female gonadal cell culture.…”

“…The spread adhered cells may be immune cells, since they had morphology similar to confluent granulocytes in C. gigas cultured heart cells [9]. Adductor muscular cells have multiple filopodia.…”

“…Many taxa in the Phylum Mollusca are marine [ 7 ]. In most cell culture studies of marine mollusks, hemocytes have been used; however, there are few reports regarding somatic cell cultures [ 8 , 9 ]. In the Pacific oyster, Crassostrea gigas , a primary culture system was developed with a variety of morphological cell types from large tissue explants [ 9 ].…”

“…In most cell culture studies of marine mollusks, hemocytes have been used; however, there are few reports regarding somatic cell cultures [ 8 , 9 ]. In the Pacific oyster, Crassostrea gigas , a primary culture system was developed with a variety of morphological cell types from large tissue explants [ 9 ]. Gonadal cell cultures were developed for this species and for the hard clam, Meretrir lusoria , with some morphological cell types from tissue explants [ 9 , 10 ].…”

“…In the Pacific oyster, Crassostrea gigas , a primary culture system was developed with a variety of morphological cell types from large tissue explants [ 9 ]. Gonadal cell cultures were developed for this species and for the hard clam, Meretrir lusoria , with some morphological cell types from tissue explants [ 9 , 10 ]. Cell cultures of digestive gland‐derived amoebocytes, adductor muscle‐derived spindle‐shaped cells, and cells with elongated protrusions were also developed from Pacific oyster tissue explants [ 9 , 10 ].…”