Development of Cell Lines from Various Tissues of Lepidoptera

Lynn, Dwight E.; Dougherty, Edward M.; McClintock, J. Thomas; Loeb, Marcia J.

doi:10.1007/978-3-642-73626-1_56

Cited by 66 publications

(46 citation statements)

References 12 publications

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“…The cell lines that we investigated were isolated from a variety of tissue types-embryonic (BTI-Tn-5b1-4, Granados et al, 1994;IPLB-LdEIta, Lynn et al, 1988), pupal ovarian (TN368, Hink, 1970), unspecified ovarian (BmN, Maeda, 1984), imaginal disks (Sf21, Vaughn et al, 1977), and larval testicular sheath (IPLB-HvT1, Lynn et al, 1988). These cell lines are composed of dedifferentiated cells.…”

Section: Discussionmentioning

confidence: 99%

N-Linked glycosylation of a baculovirus-expressed recombinant glycoprotein in insect larvae and tissue culture cells

Kulakosky¹,

Hughes²,

Wood³

1998

Glycobiology

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The potential of insect cell cultures and larvae infected with recombinant baculoviruses to produce authentic recombinant glycoproteins cloned from mammalian sources was investigated. A comparison was made of the N-linked glycans attached to secreted alkaline phosphatase (SEAP) produced in four species of insect larvae and their derived cell lines plus one additional insect cell line and larvae of one additional species. These data survey N-linked oligosaccharides produced in four families and six genera of the order Lepidoptera. Recombinant SEAP expressed by recombinant isolates of Autographa californica and Bombyx mori nucleopolyhedroviruses was purified from cell culture medium, larval hemolymph or larval homogenates by phosphate affinity chromatography. The N-linked oligosaccharides were released with PNGase-F, labeled with 8-aminonaphthalene-1-3-6-trisulfonic acid, fractionated by polyacrylamide gel electrophoresis, and analyzed by fluorescence imaging. The oligosaccharide structures were confirmed with exoglycosidase digestions. Recombinant SEAP produced in cell lines of Lymantria dispar (IPLB-LdEIta), Heliothis virescens (IPLB-HvT1), and Bombyx mori (BmN) and larvae of Spodoptera frugiperda, Trichoplusia ni, H.virescens, B.mori, and Danaus plexippus contained oligosaccharides that were structurally identical to the 10 oligosaccharides attached to SEAP produced in T.ni cell lines. The oligosaccharide structures were all mannose-terminated. Structures containing two or three mannose residues, with and without core fucosylation, constituted more than 75% of the oligosaccharides from the cell culture and larval samples.

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Section: Discussionmentioning

confidence: 99%

N-Linked glycosylation of a baculovirus-expressed recombinant glycoprotein in insect larvae and tissue culture cells

Kulakosky¹,

Hughes²,

Wood³

1998

Glycobiology

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“…Primary cultures were isolated from middevelopment H. virescens eggs obtained from Dr. James Foster (Dupont Ag, Newark, DE) using procedures described in Lynn (1996). Six primary cultures were initiated in BML-TC/10 medium (TC-100 from Sigma Chemical Co., St. Louis, MO) modified as previously described (Lynn et al, 1988) in June, 1995 and maintained at 26°C. Medium was added or re-placed on these cultures at 7-to 14-day intervals until enough cells were available for subculturing.…”

Section: Methodsmentioning

confidence: 99%

“…The stains, buffers, and gels were obtained from Innovative Chemistry (Marshfield, MA). An extract from IPLB-HvT1 cells (Lynn et al, 1988) was included on the gels for comparison and the isozyme migration patterns were also compared with other cells maintained in our laboratory.…”

Section: Characterizationmentioning

confidence: 99%

“…This insect quickly develops resistance to chemical pesticides and is a prime target for biological controls, including insect viruses (Ignoffo, 1966). While other cell lines have been established from this insect (McIntosh and Ignoffo, 1983;Lynn et al, 1988), the yields of baculovirus occlusion bodies (OBs) in these cells were lower than can be obtained in other lepidopteran lines (McIntosh et al, 1985). All of these lines were established from larval or pupal reproductive tissue.…”

Section: Introductionmentioning

confidence: 99%

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New Cell Lines fromHeliothis virescens: Characterization and Susceptibility to Baculoviruses

Lynn¹,

Shapiro²

1998

Journal of Invertebrate Pathology

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New cell lines from embryos of Heliothis virescenswere recently developed. Six primary cultures were initiated in June 1995. From these initial cultures, two produced sufficient cell growth to allow subcultivation and eventually led to the establishment of seven cell strains, three of which are maintained at low temperatures (17°C). The strains were compared with a previously established cell line from H. virescens by isozyme analysis and shown to be from the same species. All the strains were inoculated with various baculoviruses, including Autographa californica nucleopolyhedrovirus (NPV), Anagrapha falcifera NPV, Anticarsa gemmatalis NPV, Rachoplusia ou NPV, Lymantria dispar NPV (LdMNPV), Orgyia pseudotsugata NPV (OpSNPV), O. leucostigma NPV (OlMNPV), and Helicoverpa zea NPV (HzSNPV). All seven strains were highly susceptible to the noctuid NPVs, and large numbers of occlusion bodies (OBs) were produced in most of the inoculated cells. The HzSNPV infection developed at a slower rate (requiring 1 week or more before a substantial number of cells contained OBs compared with 2-3 days for the other three noctuid NPVs). Three of the H. virescens strains were also susceptible to OpSNPV although only 10-20% of the cells produced OBs with this virus. We did not observe cytopathology (CPE) in any cells inoculated with OlMNPV or LdMNPV. Our results suggest that these new strains can be useful for the study and possibly the production of baculoviruses for which no effective cell systems are available and for comparative studies on multiple virus species.

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“…In general, these lines were established from tissues of the natural host of the baculovirus to replicate, because viral yields tend to be higher in infected cultures of these homologous cell lines than in the heterologous ones. For instance, the cell lines saUFL-AG-286 (Anticarsia gemmatalis) (Sieburth & Maruniak, 1988), BM5 (Bombyx mori) (Grace, 1967), BCIRL-HZ-AM1 (Heliothis zea) (McIntosh & Ignoffo, 1981) and IPLB-LdEIta (Lymantria dispar) (Lynn et al, 1988) have been used to produce specifically the viruses AgMNPV, BmNPV, HaSNPV and LdMNPV, respectively. However, these cell lines are not as well characterized as the most widely used lines, and their technological properties (adaptation to suspension, ability to grow in serum-free culture media) are less remarkable or yet unknown.…”

Section: Insect Cell Linesmentioning

confidence: 99%

Production of Insecticidal Baculoviruses in Insect Cell Cultures: Potential and Limitations

Claus¹,

Gioria²,

Micheloud³

et al. 2012

Insecticides - Basic and Other Applications

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Development of Cell Lines from Various Tissues of Lepidoptera

Cited by 66 publications

References 12 publications

N-Linked glycosylation of a baculovirus-expressed recombinant glycoprotein in insect larvae and tissue culture cells

N-Linked glycosylation of a baculovirus-expressed recombinant glycoprotein in insect larvae and tissue culture cells

New Cell Lines fromHeliothis virescens: Characterization and Susceptibility to Baculoviruses

Production of Insecticidal Baculoviruses in Insect Cell Cultures: Potential and Limitations

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