Establishment and characterization of insect cell lines from 10 lepidopteran species

Abstract

Cell lines from selected lepidopteran species were established for the overall purpose of use in baculovirus production. A total of 36 new cell lines from 10 lepidopteran species were generated, including cell lines from a pyralid, the European corn borer, Ostrinia nubilalis, a plutellid, the diamondback moth, Plutella xylostella, as well as eight noctuids: the black cutworm, Agrotis ipsilon, the celery looper, Anagrapha falcifera, the velvetbean caterpillar, Anticarsia gemmatalis, the corn earworm, Helicoverpa zea, the tobacco budworm, Heliothis virescens, the beet armyworm, Spodoptera exigua, the fall armyworm, Spodoptera frugiperda, and the cabbage looper, Trichoplusia ni. Tissues used for cell line establishment included fat bodies, ovaries, testes, or whole embryos/larvae/pupae. All the cell lines were subcultured numerous times, characterized by isoenzyme analysis and/or deoxyribonucleic acid amplification fingerprinting using polymerase chain reaction, and stored in liquid nitrogen. Many of the cell lines were adapted to grow in serum-free medium, with cell lines from A. ipsilon and H. virescens being adapted to suspension culture using shaker flasks. The potential use for these cell lines in baculovirus production is discussed.

AcMNPV enhances infection by ThorNPV in Sf21 cells and SeMNPV in Hi5 cells

“…Insect cell lines used in this study included IPLB-SF21AE (Sf21) from Spodoptera frugiperda (Invitrogen), BTI-TN-5B1-4 (Hi5) from Trichoplusia ni (Invitrogen), SeE1 from S. exigua [21] and Spex-II A from the fat bodies of S. exigua [22]. All cells were maintained in Grace's medium (Invitrogen) supplemented with 10 % fetal bovine serum at 27°C.…”

Ecdysteroid signaling in ecdysteroid-resistant cell lines from the polyphagous noctuid pest Spodoptera exigua

“…Se4 cells (BCIRL/AMCY-SeE-CLG4), originated from embryos of the beet armyworm S. exigua (Goodman et al, 2001) were maintained at 27 C in Ex-Cell Ò 420 medium (JRH Biosciences, Hampshire, UK), supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Sigma Co., Bornem, Belgium). Se4 cell lines resistant to 20E (Se4-20E-R4) or methoxyfenozide (Se4-RH2485-R4) were grown in the presence of 100 mM of their respective ecdysteroid agonist .…”

Cell-Based Screening Systems for Insecticides

“…This condition is not always satis fi ed, as illustrated by the Se4 (derived from the lepidopteran Spodoptera exigua ; Goodman et al 2001 ) and the coleopteran BCIRL-Lepd-SL1 (derived from the Colorado potato beetle Leptinotarsa decemlineata ; Long et al 2002 ) and BRL-AG-3A (derived from the cotton boll weevil Anthonomus grandis ; Stiles et al 1992 ) cell lines, which do not express reporter genes following transfection Soin et al 2009 ) . To explore alternative means of gene transduction in these cell lines, recombinant baculoviruses were generated that have incorporated the EcRE/b.act/GFP cassette .…”