Next-generation cell lines established from the fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae)

Reall, Tamra; Kraus, Susanne; Goodman, Cynthia L.; Ringbauer, Joseph A.; Geibel, Sven; Stanley, David

doi:10.1007/s11626-019-00394-9

Cited by 18 publications

(12 citation statements)

References 21 publications

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“…Some of these lines are in routine use in industry, university, and government laboratories. The Biological Control of Insects Research Laboratory (BCIRL) has a history of establishing cell lines [63][64][65][66][67][68], generally using a standard protocol. This protocol has a core set of steps systematically repeated with observation-based changes in media components that ultimately lead to established, functional cell lines.…”

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

confidence: 99%

“…In later iterations, cell lines are initiated with other media, and sometimes with new media created by mixing known media or by adding media supplements. This iterative process generally leads to the establishment of permanent cell lines, useful in several research and development programs [65,67,68].…”

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

confidence: 99%

“…Cell cultures were maintained at 28 or 33 • C and observed daily. Insect cell lines are usually maintained at 28 • C [61, [63][64][65][66][67][68]. We chose 33 • C as a comparison temperature, because the honey bee brood nest temperature is normally maintained at 33-36 • C for larval and pupal development [70].…”

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

confidence: 99%

“…The influence of the media + FBS on overall cell health was evaluated by visual inspection before testing different combinations of basal media or comparing the effects of supplements (nutritional or hormonal) added to the media. For example, we initiated cell lines with CLG#2, a combination of an insect cell culture medium (EX-CELL 420) and a mammalian cell culture medium (L-15), used to establish lepidopteran and hemipteran cell lines [67,68]). The HZ media mixture series began with the observation that CLG#2 produced healthy bee cell cultures.…”

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

confidence: 99%

“…This slow growth rate, combined with cells that are often difficult to culture, suggests that the limiting dilution method will be more challenging for bee cells. To promote cell replication, Reall et al [68] used a conditioned medium from 72 hr old (log growth phase) parent cell lines, containing naturally produced growth factors, to generate clonal lines from S. frugiperda nervous system cell lines (7:3 conditioned medium to fresh medium). Cells were fed every 7 to 10 days with conditioned medium while in the 96-well plate, and with fresh media after they were transferred into T 12.5 flasks.…”

Section: Subcloning To Establish a Virus-free Cell Linementioning

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: A Systematic Iterative Protocol To Establish Tissue-derived mentioning

confidence: 99%

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

confidence: 99%

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

confidence: 99%

Section: Subcloning To Establish a Virus-free Cell Linementioning

confidence: 99%

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Cell Lines for Honey Bee Virus Research

Guo

Goodman

Stanley

et al. 2020

Viruses

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Prostaglandin A₂ induces apoptosis in three cell lines derived from the fall armyworm, Spodoptera frugiperda

Wang

Goodman

Ringbauer

et al. 2021

Arch Insect Biochem Physiol

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Animals maintain homeostasis of cell numbers, constantly creating new cells and eliminating others. Programmed cell death, apoptosis, is a mechanism of cell elimination and it acts in many aspects of animal biology. Drawing on the biomedical background, several signals launch the apoptosis mechanisms, including prostaglandins (PGs). Based on this information, we posed the hypothesis that PGs similarly induce apoptosis in insect cell lines. We used three Spodoptera frugiperda cell lines, including two newly established, BCIRL-SfNS-0518B-YL derived from the central nervous system and BCIRL-Sf4FB-0614-SGS derived from fat body, and the commercially available Sf9 cells. Using a kinetic apoptosis kit, we found treating SfNS cells for 18 h with 15 or 20 μM PGA 2 led to decreases in cell numbers, coupled with increased numbers of apoptotic and dead cells. Similar exposures to 10 μM PGA 2 (24 h) led to substantial increases in apoptotic cells, confirmed by a terminal deoxynucleotidyl transferase dUTP nick end labeling assay on a flow cytometer. The influence of PGA 2 treatments increased with dosage, as we recorded about 20% apoptosis at 24 h post-PGA 2 treatments (10 μM) and about 34% apoptosis at 24 h post-30 μM treatments. PGA 2 treatments led to 10-to 30-fold increases in messenger RNAs (mRNAs) encoding apoptosis-specific caspases-1, −2, −3, and −5 at 12 h and 40-to 60-fold increases in mRNAs encoding

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Indomethacin and 20‐hydroxyecdysone influence protein expression in a Spodoptera frugiperda nervous system cell line

Wang,

Buer,

Goodman

et al. 2024

Arch Insect Biochem Physiol

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Insecticide mode of action studies provide insights into how new insecticidal actives function and contribute to assessing safety to humans and nontarget organisms. Insect cell lines that express potential target sites can serve as valuable tools in this effort. In this paper, we report on the influence of two signaling molecules on protein expression in a nervous system cell line established from Spodoptera frugiperda (Bayer/BCIRL‐SfNS2‐0714‐TR). We selected this line because we established it in our laboratory and we are experienced in using it. Cells were exposed to the insect developmental hormone (1 µg/mL 20‐hydroxyecdysone, 20E) and/or a cyclooxygenase (COX) inhibitor (25 μM indomethacin, INDO; inhibits prostaglandin [PG] biosynthesis) for 24 h (Day 2), 72 h (Day 4), or 120 h (Day 6). We selected a PG biosynthesis inhibitor because PGs act in many aspects of insect biology, such as embryonic development, immunity, and protein phosphorylation. We selected the developmental hormone, 20E, because it also acts in fundamental aspects of insect biology. We identified specific proteins via in silico analysis. Changes in protein expression levels were determined using liquid chromatography‐mass spectrometry (MS) + MS‐MS. The largest number of changes in protein expression occurred on Day 2. The combination of 20E plus INDO led to 222 differentially expressed proteins, which documents the deep significance of PGs and 20E in insect biology. 20E and, separately, INDO led to changes in 30 proteins each (p value < 0.01; >2X or <0.5X‐fold changes). We recorded changes in the expression of 9 or 12 proteins (20E), 10 or 6 proteins (INDO), and 21 or 20 proteins (20E + INDO) on D4 and D6, respectively. While the cell line was established from neuronal tissue, the differentially expressed proteins act in a variety of fundamental cell processes. In this paper, we moved beyond a list of proteins by providing detailed, Gene Ontology term analyses and enrichment, which offers an in‐depth understanding of the influence of these treatments on the SfNS2 cells. Because proteins are active components of cell physiology in their roles as enzymes, receptors, elements of signaling transduction pathways, and cellular structures, changes in their expression levels under the influence of signaling molecules provide insights into their function in insect cell physiology.

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Next-generation cell lines established from the fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae)

Cited by 18 publications

References 21 publications

Cell Lines for Honey Bee Virus Research

Cell Lines for Honey Bee Virus Research

Prostaglandin A₂ induces apoptosis in three cell lines derived from the fall armyworm, Spodoptera frugiperda

Indomethacin and 20‐hydroxyecdysone influence protein expression in a Spodoptera frugiperda nervous system cell line

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Next-generation cell lines established from the fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae)

Cited by 18 publications

References 21 publications

Cell Lines for Honey Bee Virus Research

Cell Lines for Honey Bee Virus Research

Prostaglandin A2 induces apoptosis in three cell lines derived from the fall armyworm, Spodoptera frugiperda

Indomethacin and 20‐hydroxyecdysone influence protein expression in a Spodoptera frugiperda nervous system cell line

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Prostaglandin A₂ induces apoptosis in three cell lines derived from the fall armyworm, Spodoptera frugiperda