Marine Organism Cell Biology and Regulatory Sequence Discoveryin Comparative Functional Genomics

Barnes, David W.; Mattingly, Carolyn J.; Parton, Angela; Dowell, Lori; Bayne, Christopher J.; Forrest, Jeffrey Yi‐Lin

doi:10.1007/s10616-005-1719-5

Cited by 7 publications

(16 citation statements)

References 79 publications

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“…We confirmed (Bayne and Parton 2004, 2005; Barnes et al 2004) that such low concentrations of nutrient media are well suited to cell cultures of one other marine invertebrate, an echinoderm Stongylocentrotus sp. Brandt, 1835.…”

Section: Difficulties In Establishing Molluscan Cell Lines and Possisupporting

confidence: 67%

Molluscan cells in culture: primary cell cultures and cell lines

2013

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In vitro cell culture systems from molluscs have significantly contributed to our basic understanding of complex physiological processes occurring within or between tissue-specific cells, yielding information unattainable using intact animal models. In vitro cultures of neuronal cells from gastropods show how simplified cell models can inform our understanding of complex networks in intact organisms. Primary cell cultures from marine and freshwater bivalve and gastropod species are used as biomonitors for environmental contaminants, as models for gene transfer technologies, and for studies of innate immunity and neoplastic disease. Despite efforts to isolate proliferative cell lines from molluscs, the snail Biomphalaria glabrata Say, 1818 embryonic (Bge) cell line is the only existing cell line originating from any molluscan species. Taking an organ systems approach, this review summarizes efforts to establish molluscan cell cultures and describes the varied applications of primary cell cultures in research. Because of the unique status of the Bge cell line, an account is presented of the establishment of this cell line, and of how these cells have contributed to our understanding of snail host-parasite interactions. Finally, we detail the difficulties commonly encountered in efforts to establish cell lines from molluscs and discuss how these difficulties might be overcome.

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Section: Difficulties In Establishing Molluscan Cell Lines and Possisupporting

confidence: 67%

Molluscan cells in culture: primary cell cultures and cell lines

2013

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“…The optimal medium in which phagocytes remained well spread for up to three days was echinoid coelomocyte culture medium (ECCM; Table 1) and was employed for subsequent evaluations of phagocytes in culture. ECCM is based on a medium that has been used successfully to culture cells from Polian vesicles and axial organs of S. droebachiensis [33]. It was made up of a basal nutrient medium (LDF) containing Leibovitz’s L-15 Medium, Dulbecco’s Modified Eagle Medium and F-12 Nutrient Mixture.…”

Section: Resultsmentioning

confidence: 99%

“…Here we report a modified method for short-term primary cultures of phagocytes from the purple sea urchin that is based on previous coelomocyte culture methods [33]. We report cell viability for the short-term cultures and utilize our method to investigate the changing morphology of the phagocytes and their responses to lipopolysaccharide (LPS).…”

Section: Introductionmentioning

confidence: 99%

Aggregation of Sea Urchin Phagocytes Is Augmented In Vitro by Lipopolysaccharide

2013

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Development of protocols and media for culturing immune cells from marine invertebrates has not kept pace with advancements in mammalian immune cell culture, the latter having been driven by the need to understand the causes of and develop therapies for human and animal diseases. However, expansion of the aquaculture industry and the diseases that threaten these systems creates the need to develop cell and tissue culture methods for marine invertebrates. Such methods will enable us to better understand the causes of disease outbreaks and to develop means to avoid and remedy epidemics. We report a method for the short-term culture of phagocytes from the purple sea urchin, Strongylocentrotus purpuratus, by modifying an approach previously used to culture cells from another sea urchin species. The viability of cultured phagocytes from the purple sea urchin decreases from 91.6% to 57% over six days and phagocyte morphology changes from single cells to aggregates leading to the formation of syncytia-like structures. This process is accelerated in the presence of lipopolysaccharide suggesting that phagocytes are capable of detecting this molecular pattern in culture conditions. Sea urchin immune response proteins, called Sp185/333, are expressed on the surface of a subset of phagocytes and have been associated with syncytia-like structures. We evaluated their expression in cultured phagocytes to determine their possible role in cell aggregation and in the formation of syncytia-like structures. Between 0 and 3 hr, syncytia-like structures were observed in cultures when only ∼10% of the cells were positive for Sp185/333 proteins. At 24 hr, ∼90% of the nuclei were Sp185/333-positive when all of the phagocytes had aggregated into syncytia-like structures. Consequently, we conclude that the Sp185/333 proteins do not have a major role in initiating the aggregation of cultured phagocytes, however the Sp185/333 proteins are associated with the clustered nuclei within the syncytia-like structures.

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“…2), few cell biological or genomic data are available for the most primitive jawed vertebrates, the cartilaginous fish, class Chondrichthyes, largely represented by the order Elasmobranchii (3,4). This information would extend comparative data through Ͼ450 million years of evolution.…”

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confidence: 99%

RNA expression in a cartilaginous fish cell line reveals ancient 3′ noncoding regions highly conserved in vertebrates

Forest

Nishikawa

Kobayashi

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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We have established a cartilaginous fish cell line [Squalus acanthias embryo cell line (SAE)], a mesenchymal stem cell line derived from the embryo of an elasmobranch, the spiny dogfish shark S. acanthias. Elasmobranchs (sharks and rays) first appeared >400 million years ago, and existing species provide useful models for comparative vertebrate cell biology, physiology, and genomics. Comparative vertebrate genomics among evolutionarily distant organisms can provide sequence conservation information that facilitates identification of critical coding and noncoding regions. Although these genomic analyses are informative, experimental verification of functions of genomic sequences depends heavily on cell culture approaches. Using ESTs defining mRNAs derived from the SAE cell line, we identified lengthy and highly conserved gene-specific nucleotide sequences in the noncoding 3 UTRs of eight genes involved in the regulation of cell growth and proliferation. Conserved noncoding 3 mRNA regions detected by using the shark nucleotide sequences as a starting point were found in a range of other vertebrate orders, including bony fish, birds, amphibians, and mammals. Nucleotide identity of shark and human in these regions was remarkably well conserved. Our results indicate that highly conserved gene sequences dating from the appearance of jawed vertebrates and representing potential cis-regulatory elements can be identified through the use of cartilaginous fish as a baseline. Because the expression of genes in the SAE cell line was prerequisite for their identification, this cartilaginous fish culture system also provides a physiologically valid tool to test functional hypotheses on the role of these ancient conserved sequences in comparative cell biology.noncoding sequence ͉ sequence conservation ͉ Squalus acanthias E fforts toward a comprehensive compilation of cell biological, physiological, and genomic information from a variety of vertebrate species are providing a rich source of comparative data representing critical points in evolutionary divergence (1). Although available information is rapidly expanding for teleosts (bony fishes; ref.2), few cell biological or genomic data are available for the most primitive jawed vertebrates, the cartilaginous fish, class Chondrichthyes, largely represented by the order Elasmobranchii (3, 4). This information would extend comparative data through Ͼ450 million years of evolution. We have derived an embryonic mesenchymal stem cell line [Squalus acanthias embryo cell line (SAE)] from the elasmobranch S. acanthias, the spiny dogfish shark (5). This cell line from a cartilaginous fish provides a means to study the cell biology, physiology, and genomics of Chondrichthyes. Recently, the National Human Genome Research Institute targeted for comprehensive genomic sequencing Leucoraja erinacia, an elasmobranch of the superorder Squaliformes, related to S. acanthias, underscoring the value of Chondrichthyes for comparative biology (6) In efforts to explore the cell biology and genomics of c...

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Marine Organism Cell Biology and Regulatory Sequence Discoveryin Comparative Functional Genomics

Cited by 7 publications

References 79 publications

Molluscan cells in culture: primary cell cultures and cell lines

Molluscan cells in culture: primary cell cultures and cell lines

Aggregation of Sea Urchin Phagocytes Is Augmented In Vitro by Lipopolysaccharide

RNA expression in a cartilaginous fish cell line reveals ancient 3′ noncoding regions highly conserved in vertebrates

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