Department of Zoology, Goa University, Taleigao Plateau, Goa, 403206, India,
Cytotechnology. 2013 Mar;65(2):253-62. doi: 10.1007/s10616-012-9482-x. Epub 2012 Jul 25.
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.
我们使用锯缘青蟹（Scylla serrata（Forskal））睾丸细胞培养来研究白斑综合征病毒（WSSV）的影响。我们展示了蟹类细胞培养对 WSSV 的易感性。在 L15 和蟹盐补充表皮生长因子的培养基中，增殖的睾丸细胞培养物维持了超过 4 个月。用不同浓度的病毒接种的细胞培养物显示出明显的细胞病变效应，如细胞外观改变、收缩和细胞溶解。通过巢式 PCR 技术证实了细胞培养物中 WSSV 的感染。使用 10 个碱基引物生成的 RAPD 图谱显示了病毒 DNA 在培养细胞中的掺入。未暴露于 WSSV 的对照未显示出细胞病变效应。这项工作表明，增殖的睾丸细胞培养物对于使用分子工具研究 WSSV 感染非常有用。因此，由于为 WSSV 诊断和药物治疗开辟了新的前景，因此本报告具有重要意义。
