Improved primary cell culture and subculture of lymphoid organs of the greasyback shrimp Metapenaeus ensis

Han, Qi; Li, Pengtao; Lu, Xiongbin; Guo, Zijuan; Guo, Huarong

doi:10.1016/j.aquaculture.2013.06.024

Cited by 38 publications

(9 citation statements)

References 31 publications

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“…This may be due to the insufficient nutrients when compared with the 2 × L‐15 medium, which is a similar finding to those in previous studies (Han et al. ). Among the commercially available cell culture media, M199 is the second most popular medium used to develop crustacean cell lines (Maeda et al.…”

Section: Discussionsupporting

confidence: 90%

The Development and Characterization of a Cell Culture System from Indian Mud Crabs Scylla serrata

et al. 2019

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Commercially available culture media and supplements were tested for their potential to produce primary cell cultures from tissues of Indian mud crabs Scylla serrata. Eight commercially available culture media from Sigma-Aldrich (Leibovitz's L-15, Medium 199, Grace's Insect Medium, Minimal Essential Medium, Dulbecco's Modified Eagle Medium, TC-100 Insect Medium, IPL-41 Insect Medium, and Roswell Park Memorial Institute) were examined. Three different supplements (amino acid and sugar [AS], crab muscle extract [CME], and natural seawater [NSW]) were also examined. The hemocyte culture appeared to grow well for a maximum period of 21 d in 2 × L-15 medium supplemented with AS and 15% fetal bovine serum (FBS). Partial amplification and sequencing of the cytochrome oxidase subunit I (COI) gene confirmed that the primary hemocytes originated from Indian mud crabs. The effects of four metals on hemocyte viability were evaluated using the MTT assay. Of the four metals examined (arsenic, lead, cobalt, and nickel), cobalt and nickel were more toxic to the crab cells than the other metals. Both acridine orange/ethidium bromide and Hoechst staining showed the presence of apoptosis and necrosis in metal-treated groups, which suggests that metals in an aquatic environment induce death of the Indian mud crab's hemocytes. The hemocyte primary cell culture was also used to study the cytotoxicity effect of bacterial extracellular products from Vibrio harveyi and white spot syndrome virus. This study demonstrates that hemocyte primary cell culture can be used as a tool to study viral and bacterial pathogenesis and to assess the cytotoxicity of pollutants present in aquatic environments.

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Section: Discussionsupporting

confidence: 90%

The Development and Characterization of a Cell Culture System from Indian Mud Crabs Scylla serrata

et al. 2019

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“…Primary cell cultures derived from shrimp lymphoid organ have been reported in many other publications (Assavalapsakul et al, 2003;Han et al, 2013;Wang et al, 2000) and their susceptibility to WSSV has already been demonstrated (Li et al, 2014;Tapay et al, 1995). However, little is known about the different steps of the replication cycle.…”

Section: Discussionmentioning

confidence: 89%

“…Cell cultures are basic and useful tools for the study of replication cycles of viruses and the development of antivirals. Many trials have been performed on the development of cell cultures from different organs of various crustaceans (mainly shrimp) with increasing frequency over the last 30 years (Assavalapsakul et al, 2003;Han et al, 2013;Kasornchandra et al, 1999;Li et al, 2014;Wang et al, 2000). In brief, cells derived from lymphoid, ovary and haematopoietic tissues were maintained for a certain period, and some of these cell cultures were reported to be susceptible to WSSV (Jiravanichpaisal et al, 2006;Li et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

Virus replication cycle of white spot syndrome virus in secondary cell cultures from the lymphoid organ of Litopenaeus vannamei

Desmarets

Gryse

et al. 2015

Journal of General Virology

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The replication cycle of white spot syndrome virus (WSSV) was investigated in secondary cell cultures from the lymphoid organ of Litopenaeus vannamei. The secondary cells formed a confluent monolayer at 24 h post-reseeding, and this monolayer could be maintained for 10 days with a viability of 90 %. Binding of WSSV to cells reached a maximum (73¡3 % of cells and 4.84¡0.2 virus particles per virus-binding cell) at 120 min at 4 8C. WSSV entered cells by endocytosis. The co-localization of WSSV and early endosomes was observed starting from 30 min post-inoculation (p.i.). Double indirect immunofluorescence staining showed that all cell-bound WSSV particles entered these cells in the period between 0 and 60 min p.i. and that the uncoating of WSSV occurred in the same period. After 1 h inoculation at 27 8C, the WSSV nucleocapsid protein VP664 and envelope protein VP28 started to be synthesized in the cytoplasm from 1 and 3 h p.i., and were transported into nuclei from 3 and 6 h p.i., respectively. The percentage of cells that were VP664-and VP28-positive in their nuclei peaked (50¡4 %) at 12 h p.i. Quantitative PCR showed that WSSV DNA started to be synthesized from 6 h p.i. In vivo titration of the supernatants showed that the progeny WSSV were released from 12 h p.i. and peaked at 18 h p.i. In conclusion, the secondary cell cultures from the lymphoid organ were proven to be ideal for examination of the replication cycle of WSSV.

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“…The tissues were then cut into 1 mm 3 sections and placed in 24-well culture plates (Gibco), with each well containing 2 ml of M199 medium (20% FBS (fatal bovine serum), 100 units/ml penicillin, 100 μg/ml streptomycin and 50 μg/ml kanamycin; Gibco) and etoposide at various concentrations (0, 30, 60, 90, 120 and 150 μM, Novartis Pharma). Tissues were then incubated for 24 h at 26°C–27°C [ 27 – 29 ]. Each experiment designed three parallel controls.…”

Section: Methodsmentioning

confidence: 99%

FOXL2 down-regulates vitellogenin expression at mature stage in Eriocheir sinensis

et al. 2015

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Improved primary cell culture and subculture of lymphoid organs of the greasyback shrimp Metapenaeus ensis

Cited by 38 publications

References 31 publications

The Development and Characterization of a Cell Culture System from Indian Mud Crabs Scylla serrata

The Development and Characterization of a Cell Culture System from Indian Mud Crabs Scylla serrata

Virus replication cycle of white spot syndrome virus in secondary cell cultures from the lymphoid organ of Litopenaeus vannamei

FOXL2 down-regulates vitellogenin expression at mature stage in Eriocheir sinensis

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