Protocol detail

Chikungunya Virus Propagation in Aedes albopictus Cells

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The CHIKV (06.21_E1-A226V isolate, La Reunion Island) [14 (link)] strain was used in this study. The isolate was supplied by Anna-Bella Failloux and amplified on an Ae. albopictus cell line (C6/36). Briefly, C6/36 cells were grown in sterile cell culture flasks (T25, 50 capacity) and maintained in a Leibovitz‘s medium (Gibco, Thermo Fisher Scientific, Reinach, Switzerland) supplemented with 1% antibiotics-antimycotics (penicillin, streptomycin, and amphotericin B) and 4% fetal bovine serum (FBS). Confluent layers of C6/36 cells were inoculated with 100 µL of the original isolate and incubated at 28 °C w/o CO₂ for three days. After the incubation, the supernatant from the infected flask was propagated another time in a new flask layered with a C6/36 cell line generating a P2 passage.

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Veronesi E., Paslaru A., Ettlin J., Ravasi D., Flacio E., Tanadini M, & Guidi V. (2023). Estimating the Impact of Consecutive Blood Meals on Vector Competence of Aedes albopictus for Chikungunya Virus. Pathogens, 12(6), 849.

Publication 2023

Leibovitz‘s medium

Amphotericin b Antibiotics Cell culture Cell line Cells Fetal bovine serum Penicillin Sterile Strain Streptomycin

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Variable analysis

independent variables

Inoculation of C6/36 cells with 100 µL of the original CHIKV (06.21_E1-A226V isolate, La Reunion Island) strain

dependent variables

Propagation of the virus in C6/36 cell line (generation of P2 passage)

control variables

Leibovitz's medium (Gibco, Thermo Fisher Scientific, Reinach, Switzerland) supplemented with 1% antibiotics-antimycotics (penicillin, streptomycin, and amphotericin B) and 4% fetal bovine serum (FBS)
Incubation of inoculated C6/36 cells at 28 °C without CO2

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