Small intestine of neonatal pig with porcine transmissible gastroenteritis.
Characteristics
Comments
This item is derived by adaptation of virus isolated from small intestine of gnotobiotic infected pig (ATCC VR-743) to ST cells (ATCC CRL-1746) at ATCC.
Mycoplasma contamination
Not detected
Handling information
Host
ST (ATCC CRL-1746)
Complete medium
EMEM (ATCC # 30-2003) + 1 μg/ mL TPCK- treated Trypsin (USB #22725).
Temperature
37°C
Atmosphere
95% Air, 5% CO2
Incubation
Incubate infected culture for 3 days at 37°C in a humidified 5% CO2 atmosphere, until CPE are well advanced through 70-80% of the culture.
Recommendation for infection
Plant cells 24 hours in advance and infect when cultures are 90-100% confluent. Remove medium and rinse cells twice with VGM without TPCK-treated trypsin. Leave the second wash over the cells for 3 hours at 37°C in a humidified 5% CO2 atmosphere. Remove the second wash from the cells and then inoculate with a small volume of virus diluted with VGM supplemented with TPCK- treated trypsin (1 μg/ mL) to provide a MOI of about 0.1. Adsorb for 1 hour at 37°C in a humidified 5% CO2 atmosphere. End adsorption by adding VGM supplemented with 1 μg/ mL of TPCK- treated trypsin.
Handling notes
This item is derived by adaptation of virus isolated from small intestine of gnotobiotic infected pig (ATCC

VR-743?) to ST cells (ATCC

CRL-1746?) at ATCC.
Key abbreviations
°C, Degrees Celsius
CO2, Carbon dioxide
CPE, Cytopathic effect
EMEM, Eagle's Minimum Essential Medium
MOI, Multiplicity of infection
ug, Microgram
ST, Porcine fetal testis cells
TPCK, L-1-tosylamido-2-phenylethyl chloromethyl ketone
History
Depositors
LJ Saif
Type of isolate
Animal; Food & Beverage
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