Increased polyhedra production in an insect cell line following X-irradiation.
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Cell Line↗
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Cytoplasmic bridges and gap junctions in an insect cell line (Aedes albopictus).
Cell pairs of an insect cell line (Aedes albopictus, clone C6/36) were used study simultaneously the diffusional and electrical properties of intercellular junctions. Diffusion studies involved injection of fluorescent molecules into one cell of a cell pair and visual inspection of their intercellular redistribution. Electrical measurements involved a dual voltage clamp method and whole-cell recording with patch pipette. The voltage clamp protocol was aimed at examining the dependency of the junctional conductance, gj, on membrane potential, Vm. Cell pairs exhibiting a voltage-dependent gj were found to allow intercellular diffusion of Lucifer Yellow CH (molecular mass, 443 Da), but not of FITC-dextran (molecular mass, 4,400 Da). This response pattern is consistent with the presence of gap junctions in the intercellular junctions. Cell pairs showing no voltage dependence of gj were found to permit intercellular diffusion of both Lucifer Yellow CH and FITC-dextran (dextran labelled with fluorescein isothiocyanate). This behaviour is compatible with the presence of cytoplasmic bridges connecting the two adjacent cells. Hence, in culture the cells investigated express two kinds of intercellular structures, gap junctions and cytoplasmic bridges.
Aedes↗
Cell cycle analysis and synchronization of the TN-368 insect cell line.
A cell cycle analysis of the Trichoplusia ni (TN-368) insect cell line is described. By means of autoradiography and percent labeled metaphase data, the cell cycle parameters were determined to be as follows: S, 4.5 hr; G2, 8.5 hr; M, 0.5 hr; G1, 1.0 hr; the total cell time being 14.5 hr. A synchronization procedure using 50 mM thymidine in a double block procedure was used to provide a method of obtaining a large number of cells in particular cell cycle phases, especially S and G2.
Animals↗
Hormonal studies of uridine utilization in an insect cell line CP-1268 derived from the codling moth Laspeyresia pomonella.
Ecdysterone decreased cellular growth and the incorporation of uridine into RNA following 4 days of hormone exposure. This hormone did not affect uridine incorporation following short-term exposure up to 25 hours. Juvenile hormone and farnesol both significantly decreased uridine uptake and incorporation into RNA; however, uridine uptake was inhibited to a greater extent than uridine incorporation. Cyclic AMP increased the incorporation of uridine into RNA but had no demonstrable effect on the uptake process. This stimulation was not the result of cAMP degradation products. Cyclic AMP and ecdysterone together produced a significant increase in uridine incorporation into RNA. These studies demonstrate the potential utilization of insect cell lines for studying the mode of action of insect developmental hormones.
Cell Line↗
Screening of insect cell lines for the production of recombinant proteins and infectious virus in the baculovirus expression system.
Eight cell lines derived from the insects Spodoptera frugiperda, Trichoplusia ni, Mamestra brassicae, and Estigmene acrea were evaluated for recombinant beta-galactosidase and infectious virus production following infection with the baculovirus Autographa californica multiple nuclear polyhedrosis virus (AcMNPV). Production was assessed on a specific (per cell and per microgram of uninfected cellular protein) and on a volumetric (per milliliter) basis. Cell density was found to be an important factor in comparing the cell lines due to a density-dependent inhibition of specific protein and virus production that appeared to result from cell-cell contact. After infection of cells at low-density specific beta-galactosidase production per cell would drop between 3- and 6-fold in five of the eight cell lines when plated on tissue culture plates at near-confluent and confluent cell densities. The cell lines Sf 21 and Sf 9 were least sensitive to cell density. After accounting for cell density effects and differences in cell size, two cell lines, BTI Tn 5B1-4 and BTI TnM, were identified that were superior to the other cell lines, including Sf 21 and Sf 9, in beta-galactosidase production. Optimal volumetric and specific beta-galactosidase production from Tn 5B1-4 and TnM cells was 2-fold and 5-fold higher, respectively, in both cell lines than the optimal production from Sf 9 or Sf 21 cells. The Tn 5B1-4 cell line also had the highest viability of all the cell lines at 3 days postinfection and could be adapted to serum-free media.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
An insect cell line persistently infected with a baculovirus-like particle.
A persistent infection by a baculovirus-like particle was found in the established lepidopteran (Heliothis zea) cell line, IMC-HZ-1. The virus caused CPE in less than 1% of the IMC-HZ-1 cells, as measured by phase-contrast and electron microscopy. Transmission tests showed that four lepidopteran cell lines were susceptible to the persistent virus (designated as IMC-HZ-I-NOV). In inoculated TN-368 cell cultures, 90--100% infection was achieved. The ultrastructure and development of IMC-HZ-I-NOV in cell cultures were similar to known baculoviruses, and it is probable that this persistent virus is a member of the family Baculoviridae. Two lepidopterous species (Estigmene acrea and H. zea) inoculated with IMC-HZ-1-NOV by intrahemocoelic injection and/or per os feeding of larvae were not susceptible.
Cell Line↗
Replication of baculoviruses in established insect cell lines: phenomenon of attenuation. Brief review.
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Cell Line↗
Utilization of uridine for RNA synthesis in the insect cell line CP-1268 derived from the codling moth, Laspeyresia pomonella.
The utilization of [3H]-5-uridine by CP-1268 cells was studied. Uridine was rapidly transported into these cells by a concentration dependent, saturable process. Exogenous uridine rapidly equilibrated with cellular nucleotide pools and virtually all of the uridine transported into the cells was phosphorylated. Uridine incorporation into RNA was studied by continuous and pulse-labeling techniques in the prescence or absence of actinomycin D and cordycepin. These studies have shown that the pattern of unstable RNA precursor and relatively stable RNA product relationship known to exist in mammalian cells similarly exists in insect cells in vitro. This pattern varied markedly with pulse-labeling time and required the addition of RNA inhibitors to block reincorporation of intracellular labeled metabolites during the chase.
Biological Transport↗
Two very different components of messenger RNA in an insect cell line.
The messenger RNA lifetimes have been measured in a cell line derived from an invertebrate source, the mosquito Aedes albopictus. The experiments were made possible by a new technique for obtaining undegraded cytoplasmic RNA from cells with high endogenous nuclease levels. There are two components to the decay kinetics of Aedes mRNA. The major fraction of the steady state message population has a half-life of 20 hr which is, as in mammalian cells, comparable to the cell generation time. The short-lived component turns over very rapidly with a half-life estimated to be about 1.2 hr. The difference in lifetime between the short and long-lived components is about 15 fold in these cells, compared to 3-4 fold in mammalian cells. This may reflect the need for a more responsive mRNA regulating system in poikilothermic organisms. The great disparity between the principle messenger lifetimes permits a more definite assignment of a two component behavior to message decay. The data in the case of mammalian cells could not rule out a family of intermediate lifetimes. The long-lived mRNA has a much smaller average sedimentation value than the short-lived material. The effect is similar to, but much larger than, that seen in mammalian cells. Although the lifetime difference is much greater in the insect cells than in human (HeLa) cells, the fast and slow components comprise about the same proportion of the steady state mRNA population: 30 percent and 70 percent, respectively.
Aedes↗
Development of an attached strain from a continuous insect cell line.
A continuous attached cell strain has been developed from the IPRI-CF-124 line of the spruce budworm, Choristoneura fumiferana. This was done by discarding suspended cells at each passage, rinsing attached cells with 0.05% trypsin and using only the strongly attached cells for subculturing. The method is very effective in that the proportion of attached cells increased from 6% in the parent cell line to 97% in the new cell strain after 20 passages. The attachment and growth properties are stable after storage of cells in liquid nitrogen. The new cell strain is designated IPRI-CF-124T and has a population doubling time comparable to that of the parent cell line.
Cell Adhesion↗
Expression of three recombinant proteins using baculovirus vectors in 23 insect cell lines.
Recombinant Autographa california baculoviruses expressing genes for pseudorabies virus glycoprotein (gp50T), human plasminogen (HPg), and beta-galactosidase (beta-gal) were used to infect 23 cell lines or strains. The objectives were to compare amounts of recombinant proteins expressed in the cell lines, compare yields from clones and parent lines, investigate the effects of long-term culture in serum-free medium on production, and determine if some lines yield gp50T with different glycosylation patterns. For HPg, IZD-MB0503 had the highest yield and four other lines (IPLB-TN-R2, IPLB-SF-1254, IPLB-LdEIta, and CM-1) had levels above that of SF-9 cells. For gp50T, four lines (IPLB-HvT1, IPLB-SF21AE, IPLB-SF21AE-15, and IPLB-SF-1254) had higher amounts than SF-9 cells. Some lines yielded gp50T with molecular mass about 1000 daltons larger than that from SF-9 cells, which suggests increased oligosaccharide processing. Equally high levels of beta-gal were expressed in three lines (SF-9, IZD-MB0503, and BCIRL-PX2-HNV3). The major conclusion is that no single cell line produced highest yields for all three recombinant proteins. Four lines were cultured in serum-free medium for 31-34 passages and then infected with the three recombinant viruses. For most cell line-recombinant combinations, the yields in serum-free medium were equal to or better than those in serum-supplemented medium. Medium composition had a much stronger effect on foreign gene expression than on susceptibility of cells to wild-type virus.
Animals↗
Effects of x-irradiation on cell division, oxygen consumption, and growth medium pH of an insect cell line cultured in vitro.
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Animals↗
[Comparison of enzyme activity in two insect cell lines: Anthera eucalypti and Malacosoma disstria (Lepidoptera)].
A new micromethod has been used to compare the enzymic profiles of two cell lines of Lepidoptera (Antheraea eucalypti and Malacosoma disstria). Differences are observed on alkaline phosphatase, on two aminopeptidases, on alpha and beta galactosidases and on alpha mannosidase. The whole intracellular activity of the enzymes revealed is detected without any previous treatment of the cells.
Alkaline Phosphatase↗
Advancing insect research through cell line transcriptomics.
This review emphasizes the significance of insect cell lines in transcriptomic research, highlighting their role as vital tools for uncovering cellular and molecular mechanisms of insect physiology, immune responses, and adaptation to environmental stressors. Cell lines derived from tissues such as the midgut, fat body, nervous system, and reproductive organs enable researchers to examine gene expression changes in a controlled setting, making discoveries that are difficult to achieve through whole-organism studies. High-throughput sequencing and single-cell RNA sequencing (scRNA-seq) have identified genes linked to detoxification, stress response, development, and immune defense, offering valuable insights for future applications in agriculture, pest control, and biotechnology. To organize this information clearly, we have summarized key findings in a table, providing an accessible overview of each cell line's important roles in transcriptomic research. This method not only highlights the adaptability of insect cell lines in functional genomics but also underscores their usefulness as model systems in pest management, virology, and bioengineering. Through utilizing transcriptomics, insect cell lines continue to advance our understanding of insect biology and foster the development of innovative strategies for sustainable crop protection and biotechnological use.
Animals↗
Production of recombinant proteins by baculovirus-infected gypsy moth cells.
An experimental study was undertaken to evaluate alternative insect cell lines to Sf9 [from Spodoptera frugiperda (fall armyworm)] for the production of recombinant proteins. Insect cell lines from two different organisms were considered: IPLB-LdEIta (LdEIta) from Lymantria dispar (gypsy moth) and IPLB-HvT1 (HvT1) from Heliothis virescens (tobacco budworm). Both LdEIta and HvT1 produced higher total activity levels of recombinant beta-galactosidase in monolayer culture than Sf9 after infection with the Autographa californica nuclear polyhedrosis virus (AcMNPV). However, only LdEIta generated a product yield (activity per milligram of total protein) which exceeded that of Sf9 (by 25%), so its growth and production characteristics were investigated in depth. LdEIta generated production levels and yields of a recombinant rotaviral protein, VP4, which exceeded those of Sf9 by 84 and 38%, respectively. In suspension culture, the LdEIta cells grew as aggregates with a doubling time several hours longer than Sf9, but the recombinant product yields of LdEIta were still higher than Sf9 by 38% in this culture environment. beta-Galactosidase expression rates and cell death rates suggested that the difference in productivity between the two hosts was due to the ability of LdEIta to survive the baculovirus infection and produce recombinant proteins longer than Sf9. The presence of LdEIta aggregates in suspension culture may be used as a method to separate live cells from dead cells, labile product, and spent medium in recombinant protein production processes.
Animals↗
Protective effect of methylcellulose and other polymers on insect cells subjected to laminar shear stress.
The relative sensitivity of two insect cell lines to laminar shear stress was determined, and the protective effect of polymers added to the growth media of two insect cell lines, Trichoplusia ni (TN-368) and Spodoptera frugiperda (SF-9), was evaluated. TN-368 and SF-9 cells were found to be equally sensitive to laminar shear stress. Methylcellulose [0.5% (w/v) Dow E4M Methocel] and dextran [4.5% (w/v)] increased the resistance of suspended cells to lysis due to laminar shear stress by factors of up to 76 and 28, respectively, compared to cells in media without additives. It was observed that the protective effect of Pluronic F-68 was concentration-dependent: 0.2% and 0.3% (w/v) F-68 increased the resistance of SF-9 cells to shear stress by factors of 15 and 42, respectively. However, increasing the concentration to 0.5% did not significantly increase the cells' resistance compared to 0.3% (w/v). F-68 at 0.2% only increased the resistance of TN-368 cells by a factor of 6. It is believed that the protection is a result of the polymer adsorbing to the cell membrane. None of the polymer additives tested had a significant effect on SF-9 or TN-368 growth rate.
Animals↗
The establishment of two cell lines from the insect Spodoptera frugiperda (Lepidoptera; Noctuidae).
The history and characteristics of two cells lines developed from primary explants of pupal tissue from the insect, Spodoptera frugiperda (J.E. Smith), are described. One cell line, IPLB-SF 21, was developed with hemolymph-supplemented medium and has been maintained continuously on the medium. The second cell line, IPLB-SF-1254, was developed with a medium containing a combination of vertebrate sera plus hemolymph and was adapted to hemolyphn-free medium at the 6th passage. The IPLB-SF-21 cell line has a population doubling time of 26 to 30 hr; the doubling time of the IPLB-SF-1254 line is 36 hr. The chromosomal morphology and distribution was typical of other lepidopteran cell lines. Serological studies showed that both cell lines have at least one antigen which also is common is tissue antigens from pupae of Spodoptera frugiperda.
Animals↗
[Experimental infection of an invertebrate cell line with a mollicute-like procaryote inducing the "lethargy of coleoptera" (author's transl)].
In vitro multiplication of a pathogenic intravacuolar mollicute-like procaryote from Melolontha melolontha L. was experimentally obtained in an insect cell line. The elongated and pleomorphic forms observed in the insect-host are reproduced in cell cultures. A third peculiar giant form is missing, showing that it does not play any role in the multiplication of the germ. The intrinsic potentialities of the germ are maintained during the successive passages, as proved by reinfection of the insect and by immunology. The original syndrome including the giant form is reproduced in the insect. The immunserum prepared from the wild germ isolated by density gradient is positive with the in vitro mollicute. The germs are intravacuolar, both in the cultured cells and in the insect host. Clearly the microorganism multiplies within the vacuoles. A cytopathogenic effect is noticed in the cultured cells overcrowed with germs. The germs become extracellular when they are released in the culture medium by disaggregation of the cell membranes. It seems that this work shows the first model of an intravacuolar mollicute-like procaryote experimentally multiplied in cultivated cells.
Animals↗