A Review of Insect Cell Culture: Establishment, Maintenance and Applications in Entomological Research

Arunkarthick, S.; Asokan, R.; Aravintharaj, R.; Niveditha, M.; Kumar, N. K. Krishna

doi:10.18474/jes17-02pt.1

Cited by 18 publications

(14 citation statements)

References 68 publications

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“…Hundreds of insect cell lines have been established since the first ones were produced in the late 1950s and 1960s [61,62]. Some of these lines are in routine use in industry, university, and government laboratories.…”

Section: A Systematic Iterative Protocol To Establish Tissue-derived mentioning

confidence: 99%

“…A nodavirus, named " Tn-nodavirus", was discovered in the BTI-TN-5B1-4 (Tn5) cell line derived from Trichoplusia ni, [86] and subsequently in a wide range of T. ni cell lines [80]. The IPLB-Sf21 cell line derived from Spodoptera frugiperda pupal ovaries, along with the subclonal line, Sf9, are well recognized for generating recombinant proteins via the baculovirus expression system [61]. These Sf cell lines are infected with the Sf-rhabdovirus [81,87].…”

Section: Use Of Antiviral Drugs To Establish Virus-free Insect Cell Lmentioning

confidence: 99%

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Cell Lines for Honey Bee Virus Research

Guo

Goodman

Stanley

et al. 2020

Viruses

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With ongoing colony losses driven in part by the Varroa mite and the associated exacerbation of the virus load, there is an urgent need to protect honey bees (Apis mellifera) from fatal levels of virus infection and from the non-target effects of insecticides used in agricultural settings. A continuously replicating cell line derived from the honey bee would provide a valuable tool for the study of molecular mechanisms of virus-host interaction, for the screening of antiviral agents for potential use within the hive, and for the assessment of the risk of current and candidate insecticides to the honey bee. However, the establishment of a continuously replicating honey bee cell line has proved challenging. Here, we provide an overview of attempts to establish primary and continuously replicating hymenopteran cell lines, methods (including recent results) of establishing honey bee cell lines, challenges associated with the presence of latent viruses (especially Deformed wing virus) in established cell lines and methods to establish virus-free cell lines. We also describe the potential use of honey bee cell lines in conjunction with infectious clones of honey bee viruses for examination of fundamental virology.

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Section: A Systematic Iterative Protocol To Establish Tissue-derived mentioning

confidence: 99%

Section: Use Of Antiviral Drugs To Establish Virus-free Insect Cell Lmentioning

confidence: 99%

Cell Lines for Honey Bee Virus Research

Guo

Goodman

Stanley

et al. 2020

Viruses

View full text Add to dashboard Cite

show abstract

“…A nodavirus, named " Tn-nodavirus", was discovered in the BTI-TN-5B1-4 (Tn5) cell line derived from Trichoplusia ni, [86] and subsequently in a wide range of T. ni cell lines [80]. The IPLB-Sf21 cell line derived from Spodoptera frugiperda pupal ovaries, along with the subclonal line, Sf9, are well-recognized for generating recombinant proteins via the baculovirus expression system [87].…”

Section: Use Of Antiviral Drugs To Establish Virus-free Insect Cell Lmentioning

confidence: 99%

“…Cell cultures are maintained at 28 or 33 o C and observed daily. Insect cell lines are usually maintained at 28 o C[61,[63][64][65][66][67][68]. We chose 33 o C as a comparison temperature because the honey bee brood nest temperature is maintained at 33-36 o C for larval and pupal development[70].Cultures are fed every 4 to 14 days (either by adding medium or replacing half, with these final concentrations of antibiotics: 50 U/mL penicillin, 0.05 mg/mL streptomycin).Over 600 honey bee cell cultures have been initiated using various combinations of tissues, media and media additives(Table 3).…”

mentioning

confidence: 99%

Cell Lines for Honey Bee Virus Research

Guo¹,

Goodman²,

Stanley³

et al. 2020

Preprint

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With ongoing colony losses driven in part by the Varroa mite and the associated exacerbation of virus load, there is an urgent need to protect honey bees (Apis mellifera) from fatal levels of virus infection and from nontarget effects of insecticides used in agricultural settings. A continuously replicating cell line derived from the honey bee would provide a valuable tool for study of molecular mechanisms of virus – host interaction, for screening of antiviral agents for potential use within the hive, and for assessment of the risk of current and candidate insecticides to the honey bee. However, the establishment of a continuously replicating, honey bee cell line has proved challenging. Here we provide an overview of attempts to establish primary and continuously replicating hymenopteran cell lines, methods (including recent results) for establishing honey bee cell lines, challenges associated with the presence of latent viruses (especially Deformed wing virus), in established cell lines and methods to establish virus-free cell lines. We also describe the potential use of honey bee cell lines in conjunction with infectious clones of honey bee viruses for examination of fundamental virology.

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“…Efforts to produce insect cell lines have resulted in the establishment of more than 1000 continuous cell lines from various insect species [ 1 ]. These cell lines are utilized for a broad range of biological research areas and the industrial mass production of recombinant proteins [ 2 – 4 ]. Nevertheless, cultured insect cells suitable for a specific research objective are not always available.…”

Section: Introductionmentioning

confidence: 99%

A simple method for ex vivo honey bee cell culture capable of in vitro gene expression analysis

et al. 2021

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Cultured cells are a very powerful tool for investigating biological events in vitro; therefore, cell lines have been established not only in model insect species, but also in non-model species. However, there are few reports on the establishment of stable cell lines and development of systems to introduce genes into the cultured cells of the honey bee (Apis mellifera). We describe a simple ex vivo cell culture system for the honey bee. Hemocyte cells obtained from third and fourth instar larvae were cultured in commercial Grace’s insect medium or MGM-450 insect medium for more than two weeks maintaining a normal morphology without deterioration. After an expression plasmid vector bearing the enhanced green fluorescent protein (egfp) gene driven by the immediate early 2 (IE2) viral promoter was transfected into cells, EGFP fluorescence was detected in cells for more than one week from one day after transfection. Furthermore, double-stranded RNA corresponding to a part of the egfp gene was successfully introduced into cells and interfered with egfp gene expression. A convenient and reproducible method for an ex vivo cell culture that is fully practicable for gene expression assays was established for the honey bee.

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A Review of Insect Cell Culture: Establishment, Maintenance and Applications in Entomological Research

Cited by 18 publications

References 68 publications

Cell Lines for Honey Bee Virus Research

Cell Lines for Honey Bee Virus Research

Cell Lines for Honey Bee Virus Research

A simple method for ex vivo honey bee cell culture capable of in vitro gene expression analysis

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