Anticarsia gemmatalis Cell Line Protocol

The insect cells UFL-Ag-286 [43 (link)] were grown at 27 °C in GRACE’s medium (Invitrogen, Carlsbad, CA, USA) containing 10% v/v fetal bovine serum (FBS; GBO, Kremsmünster, Austria) and supplied with antibiotics and antimycotics (Invitrogen). Initial stocks of AgMNPV-2D [44 (link),45 (link)] were multiplied in Anticarsia gemmatalis (third instar larvae) by per os infection using OBs, and by the exposition of monolayers of UFL-Ag-286 cells with BVs in plastic tissue culture flasks. The virus stocks used in all experiments were tittered by plaque assay [46 (link)] in UFL-Ag-286 cells and maintained as culture supernatants.

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Miele S.A., Cerrudo C.S., Parsza C.N., Nugnes M.V., Mengual Gómez D.L., Belaich M.N, & Ghiringhelli P.D. (2019). Identification of Multiple Replication Stages and Origins in the Nucleopolyhedrovirus of Anticarsia gemmatalis. Viruses, 11(7), 648.

Publication 2019

GRACE’s medium antibiotics and antimycotics

Antibiotics Assay Cells Infection Insect Larvae Plaque Tissue Virus

Corresponding Organization : National University of Quilmes

Other organizations : Université Paris-Saclay, CEA Paris-Saclay, Commissariat à l'Énergie Atomique et aux Énergies Alternatives, Centre National de la Recherche Scientifique

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Variable analysis

independent variables

Temperature (27 °C)
Growth medium (GRACE's medium)

dependent variables

Growth of UFL-Ag-286 cells
Multiplication of AgMNPV-2D virus stocks

control variables

Fetal bovine serum (10% v/v)
Antibiotics and antimycotics (Invitrogen)

controls

Positive control: Multiplying AgMNPV-2D virus stocks in Anticarsia gemmatalis (third instar larvae)
Negative control: Not mentioned

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