Propagation and Plaque Purification of SADS-CoV

IPEC-J2 porcine epithelial cells were propagated in Dulbecco's modified eagle medium (D MEM) (Gibco, USA) supplemented with 10% fetal bovine serum (FBS), 1% penicillin/streptomycin (Gibco), 1% insulin-transferrin-sodium selenite (Roche), and 5 ng/ml of human epidermal growth factor (Invitrogen). The Vero (ATCC CCL-81) cells were maintained in minimal essential medium (MEM) (Gibco) supplemented with 10% heat-inactivated fetal bovine serum, 5% L-glutamine, 100 U/ml of penicillin G, and 100 μg/ml streptomycin at 37°C in a humidified 5% CO₂ incubator (27 (link)). The SADS-CoV CH/FJWT/2018 (passage 10) virus was plaque purified for three times in Vero 81 cells supplemented with a final concentration of 10 μg/ml trypsin in D MEM as described in our previous study (28 (link)).

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Zhang F., Yuan W., Li Z., Zhang Y., Ye Y., Li K., Ding Z., Chen Y., Cheng T., Wu Q., Tang Y, & Song D. (2020). RNA-Seq-Based Whole Transcriptome Analysis of IPEC-J2 Cells During Swine Acute Diarrhea Syndrome Coronavirus Infection. Frontiers in Veterinary Science, 7, 492.

Publication 2020

DMEM penicillin/streptomycin insulin-transferrin-sodium selenite human epidermal growth factor MEM penicillin G streptomycin

Cells Eagle Epidermal growth factor Epithelial cells Fetal bovine serum Glutamine Human Insulin Penicillin Penicillin g Plaque Porcine Sads cov Sodium selenite Streptomycin Transferrin Trypsin Vero cells Virus

Corresponding Organization : Jiangxi Agricultural University

Other organizations : Animal Science Research Institute, Jiangxi Academy of Agricultural Sciences

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Variable analysis

independent variables

Passage number of the SADS-CoV CH/FJWT/2018 virus

dependent variables

Growth and propagation of IPEC-J2 porcine epithelial cells
Growth and maintenance of Vero (ATCC CCL-81) cells
Plaque purification of the SADS-CoV CH/FJWT/2018 virus

control variables

Culture media for IPEC-J2 cells (DMEM supplemented with 10% FBS, 1% penicillin/streptomycin, 1% insulin-transferrin-sodium selenite, and 5 ng/ml of human epidermal growth factor)
Culture media for Vero cells (MEM supplemented with 10% heat-inactivated FBS, 5% L-glutamine, 100 U/ml of penicillin G, and 100 μg/ml streptomycin)
Culture conditions (37°C, 5% CO2 incubator)

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

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