Primary culture of the hyaline haemocytes from marine decapods

Walton, Alison M.; Smith, Valerie J.

doi:10.1006/fsim.1998.0184

Cited by 39 publications

(29 citation statements)

References 24 publications

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“…To stimulate cell growth and survival L-15 medium was supplemented with: (1) inactivated standard Fetal Calf Serum (FCS) or (2) a ''sea urchin Pluteus Estract'' (PE) specifically developed in our lab. In FCS-supplemented cell cultures signs of degeneration on were observed early and cell viability was significantly lower than in unsupplemented cultures (One-way ANOVA: P \ 0.05), suggesting that for sea urchin gonad cells this extract does not improve cell conditions, as, on the contrary, was reported for other marine invertebrates (Mulford and Austin 1998;Walton and Smith 1999;Odintsova et al 2005). Different concentrations (up to 50 lg/mL) of PE were tested but no improvement in cell conditions was obtained as well.…”

Section: Introductionmentioning

confidence: 80%

Proceedings of the symposium ‘marine invertebrate cell culture’

Costil

2013

Cytotechnology

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Section: Introductionmentioning

confidence: 80%

Proceedings of the symposium ‘marine invertebrate cell culture’

Costil

2013

Cytotechnology

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“…Primary cell cultures from different shrimp tissues such as lymphoid organ (Chen and Kou 1989;Chen and Wang 1999;Wang et al 2000), heart (Chen and Wang 1999; Owens and Smith 1999), gut (Nadala et al 1993), ovaries (Maeda et al 2004) have been used to study various viral infections and replications but long surviving and proliferating cell cultures were not then available for evaluating viral infections. Though, few researchers have attempted primary cell cultures of crabs (Ballard et al 1993;Walton and Smith 1999;Sashikumar and Desai 2008;Zeng et al 2009) none have used crabs' cell culture for studying WSSV pathogenicity.…”

Section: Discussionmentioning

confidence: 99%

Susceptibility of testicular cell cultures of crab, Scylla serrata (Forskal) to white spot syndrome virus

Shashikumar

Desai

2012

Cytotechnology

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Testicular cell culture of crab, Scylla serrata (Forskal) was used to study the effects of White spot syndrome virus (WSSV). We are showing the susceptibility of cell culture of crabs to WSSV. The proliferating cell culture of testes were maintained for more than 4 months in a medium prepared from L15 and crab saline supplemented with epidermal growth factor. The cell cultures inoculated with different concentrations of virus showed distinct cytopathic effects such as change in cell appearance, shrinkage and cell lysis. WSSV infection of cultured cells was confirmed by Nested PCR technique. The incorporation of viral DNA in cultured cells was shown by RAPD profile generated using 10-mer primers. The controls that were not exposed to WSSV did not show cytopathic effects. This work shows the usefulness of proliferating testicular cell culture for studying WSSV infection using molecular tools. Thus, this report gains significance as it opens new vistas for diagnostics and drugs for WSSV.

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“…Leudeman (1990) and Frerichs (1996) suggested that serum could be toxic for crustacean cells. Moreover Walton and Smith (1999) reported occasional clumping of cultured hemocytes of crabs (Liocarcinus depurator and Carcinus maenas) in 20% (v/v) serum, while 10% serum significantly promoted the viability of culture. Perhaps this could explain why the higher level of sera i.e.…”

Section: Discussionmentioning

confidence: 99%

Development of cell line from the testicular tissues of crab Scylla serrata

Shashikumar

Desai

2011

Cytotechnology

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This is the first report on development of a finite cell line from testicular tissues of crab, Scylla serrata. Both the explant and segregated tissues of testes yielded cells that could proliferate and grow. These cells ranged in size from 10 to 38 lm with distinct nuclei of varying shapes. The testicular cells survived and proliferated best in L-15-crab saline medium supplemented with epidermal growth factor (20 ng/mL) and glucose (1 mg/mL). The cell proliferation rate was assessed by Methyl tetrazolium assay in terms of change in optical density which clearly indicated a prominent increase in cell density. The testicular cells were subcultured at an interval of 4-6 days. These subcultured cells remained healthy and proliferated for 5 months with a minimum of ten subsequent passages. The finite cell line was characterized in terms of morphology, growth rate, lactate dehydrogenase release (for detecting health status) and 18S rRNA sequencing. This cell line could be a very useful tool for testing infections and replications of crustacean viruses. The present work provides a technique that could be extended for developing other crustacean cell lines.

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Primary culture of the hyaline haemocytes from marine decapods

Cited by 39 publications

References 24 publications

Proceedings of the symposium ‘marine invertebrate cell culture’

Proceedings of the symposium ‘marine invertebrate cell culture’

Susceptibility of testicular cell cultures of crab, Scylla serrata (Forskal) to white spot syndrome virus

Development of cell line from the testicular tissues of crab Scylla serrata

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