Cultivation and Propagation of Diverse Cell Lines and Viruses

PK-15, MARC-145, 293T, and COS-7 cells were purchased from China Center for Type Culture Collection (CCTCC, Wuhan, China). MARC-145 and 293T cells were grown in Dulbecco’s minimal essential medium (DMEM) supplemented with 10% fetal bovine serum FBS (Gibco, Carlsbad, CA, USA). PK-15 and COS-7 cells were grown in minimal essential medium (MEM) supplemented with 10% FBS. PAMs were collected from PRRSV negative pigs (28 days old) and maintained in RPMI-1640 medium supplemented with 10% FBS. Wild-type highly pathogenic PRRSV (HP-PRRSV) SD16 strain was isolated by plaque purification and grown in MARC-145 cells as previously described53 (link). HP-PRRSV SD16-EGFP recombinant virus was engineered as described previously33 (link). Pig rotavirus (AV-55) was purchased from China Center of Veterinary Culture Collection and was propagated on MA-104 cells54 .

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Gao J., Xiao S., Xiao Y., Wang X., Zhang C., Zhao Q., Nan Y., Huang B., Liu H., Liu N., Lv J., Du T., Sun Y., Mu Y., Wang G., Syed S.F., Zhang G., Hiscox J.A., Goodfellow I, & Zhou E.M. (2016). MYH9 is an Essential Factor for Porcine Reproductive and Respiratory Syndrome Virus Infection. Scientific Reports, 6, 25120.

Publication 2016

PK-15 MARC-145 COS-7 fetal bovine serum FBS

293t cells Cells Cos 7 cells Marc Pathogenic Plaque Prrsv Rotavirus Strain Virus

Corresponding Organization :

Other organizations : Northwest A&F University, Shandong Agricultural University, Harbin Veterinary Research Institute, Henan Agricultural University, University of Liverpool, Addenbrooke's Hospital, University of Cambridge

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Protocol cited in 3 other protocols

Variable analysis

independent variables

PK-15 cells
MARC-145 cells
293T cells
COS-7 cells
Wild-type highly pathogenic PRRSV (HP-PRRSV) SD16 strain
HP-PRRSV SD16-EGFP recombinant virus
Pig rotavirus (AV-55)

dependent variables

Growth of MARC-145 and 293T cells in DMEM with 10% FBS
Growth of PK-15 and COS-7 cells in MEM with 10% FBS
Growth of PAMs in RPMI-1640 with 10% FBS
Propagation of HP-PRRSV SD16 strain in MARC-145 cells
Propagation of HP-PRRSV SD16-EGFP recombinant virus
Propagation of pig rotavirus (AV-55) in MA-104 cells

control variables

PRRSV negative pigs (28 days old) for PAMs collection

positive controls

None specified

negative controls

None specified

Annotations

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