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

ABSTRACT: 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 ?m 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.

SUBMITTER: Shashikumar A

PROVIDER: S-EPMC3176930 | biostudies-literature | 2011 Oct

REPOSITORIES: biostudies-literature

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

Shashikumar Anumol A Desai P V PV

Cytotechnology 20110729 5

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 μm 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 te ...[more]

PMID: 21800082

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Project description:The giant mud crab Scylla serrata provides an important source of income and food to coastal communities in East Africa. However, increasing demand and exploitation due to the growing coastal population, export trade, and tourism industry are threatening the sustainability of the wild stock of this species. Because effective management requires a clear understanding of the connectivity among populations, this study was conducted to assess the genetic diversity and connectivity in the East African mangrove crab S. serrata. A section of 535 base pairs of the cytochrome oxidase subunit I (COI) gene and eight microsatellite loci were analysed from 230 tissue samples of giant mud crabs collected from Kenya, Tanzania, Mozambique, Madagascar, and South Africa. Microsatellite genetic diversity (He) ranged between 0.56 and 0.6. The COI sequences showed 57 different haplotypes associated with low nucleotide diversity (current nucleotide diversity = 0.29%). In addition, the current nucleotide diversity was lower than the historical nucleotide diversity, indicating overexploitation or historical bottlenecks in the recent history of the studied population. Considering that the coastal population is growing rapidly, East African countries should promote sustainable fishing practices and sustainable use of mangrove resources to protect mud crabs and other marine fauna from the increasing pressure of exploitation. While microsatellite loci did not show significant genetic differentiation (p > 0.05), COI sequences revealed significant genetic divergence between sites on the East coast of Madagascar (ECM) and sites on the West coast of Madagascar, mainland East Africa, as well as the Seychelles. Since East African countries agreed to achieve the Convention on Biological Diversity (CBD) target to protect over 10% of their marine areas by 2020, the observed pattern of connectivity and the measured genetic diversity can serve to provide useful information for designing networks of marine protected areas.

Project description:BackgroundThe fishery and aquaculture of the widely distributed mangrove crab Scylla serrata is a steadily growing, high-value, global industry. Climate change poses a risk to this industry as temperature elevations are expected to threaten the mangrove crab habitat and the supply of mangrove crab juveniles from the wild. It is therefore important to understand the genomic and molecular basis of how mangrove crab populations from sites with different climate profiles respond to heat stress. Towards this, we performed RNA-seq on the gill tissue of S. serrata individuals sampled from 3 sites (Cagayan, Bicol, and Bataan) in the Philippines, under normal and heat-stressed conditions. To compare the transcriptome expression profiles, we designed a 2-factor generalized linear model containing interaction terms, which allowed us to simultaneously analyze within-site response to heat-stress and across-site differences in the response.ResultsWe present the first ever transcriptome assembly of S. serrata obtained from a data set containing 66 Gbases of cleaned RNA-seq reads. With lowly-expressed and short contigs excluded, the assembly contains roughly 17,000 genes with an N50 length of 2,366 bp. Our assembly contains many almost full-length transcripts - 5229 shrimp and 3049 fruit fly proteins have alignments that cover >80% of their sequence lengths to a contig. Differential expression analysis found population-specific differences in heat-stress response. Within-site analysis of heat-stress response showed 177, 755, and 221 differentially expressed (DE) genes in the Cagayan, Bataan, and Bicol group, respectively. Across-site analysis showed that between Cagayan and Bataan, there were 389 genes associated with 48 signaling and stress-response pathways, for which there was an effect of site in the response to heat; and between Cagayan and Bicol, there were 101 such genes affecting 8 pathways.ConclusionIn light of previous work on climate profiling and on population genetics of marine species in the Philippines, our findings suggest that the variation in thermal response among populations might be derived from acclimatory plasticity due to pre-exposure to extreme temperature variations or from population structure shaped by connectivity which leads to adaptive genetic differences among populations.

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

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

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