virus production-serum free medium vp-sfm  (Thermo Fisher)

Journal: Journal of Microbiology (Seoul, Korea)

Article Title: CA-CAS-01-A: A Permissive Cell Line for Isolation and Live Attenuated Vaccine Development Against African Swine Fever Virus

doi: 10.1007/s12275-024-00116-1

Figure Lengend Snippet: Adaptation of ASFV in CAS-01 cells. The ASFV/INJE/11893/2021 or ASFV/INJE/13167/2021 virus was infected with CAS-01 cells at 1 × 10 5 cells/flask in a T25 flask and incubated in 2% FBS and 1% P/S added VP-SFM. After 24 h, media was changed by VP-SFM added 2% FBS, and incubated 6 days in a 5% CO 2 atmosphere at 37 °C. A – D ASFV/INJE/11893/2021, E – G ASFV/INJE/13167/2021 infection results are shown. A Immunocytochemistry (ICC) analysis using immunoperoxidase assay with ASFV p30 antibody for indicated ASFV passages to detect the ASFV infectivity. B , E Ct value was measured on the 7 dpi for each passage. The replicated viruses were titrated using C , F HAD 50 and D , G TCID 50 in the indicated ASFV passages

Article Snippet: Next, isolated ASFV/INJE/11893 and ASFV/INJE/13167/2021 was infected into cells separately, and culture media was replaced with virus production-serum free medium (VP-SFM) (Gibco™).

Techniques: Virus, Infection, Incubation, Immunocytochemistry

Journal: Journal of Microbiology (Seoul, Korea)

Article Title: CA-CAS-01-A: A Permissive Cell Line for Isolation and Live Attenuated Vaccine Development Against African Swine Fever Virus

doi: 10.1007/s12275-024-00116-1

Figure Lengend Snippet: Growth kinetics of ASFV in Primary PAM and CAS-01 cell. ASFV/INJE/11893/2021 was serially passaged twelve times in CAS-01 cells. Primary PAM ( A , D , G ) and CAS-01 cells ( B , C , E , F , H , I ) were incubated at 5 × 10 5 cells/well or 1 × 10 5 cells/well, respectively in a 6 well plate at 37 °C and 5% CO 2 atmosphere. After 12 h of cell incubation, cell passaged P-2 or P-12 ASFV was infected using VP-SFM with 1% FBS and 1% S/P. Cells were harvested at 12-h intervals up to 144 h after virus infection. After freezing and thawing (F/T) twice, Ct values A – C were obtained using RT-PCR. Viruses were titrated using D – F 50% hemadsorption doses (HAD 50 ) and G – I 50% tissue culture infectious dose (TCID 50 )

Article Snippet: Next, isolated ASFV/INJE/11893 and ASFV/INJE/13167/2021 was infected into cells separately, and culture media was replaced with virus production-serum free medium (VP-SFM) (Gibco™).

Techniques: Incubation, Infection, Virus, Reverse Transcription Polymerase Chain Reaction