Three- to four-week-old male golden Syrian hamsters (HsdHan:AURA strain) were purchased from Envigo. All studies were conducted under a protocol approved by the UTMB Institutional Animal Care and Use Committee and complied with USDA guidelines in a laboratory accredited by the Association for Assessment and Accreditation of Laboratory Animal Care. Procedures involving infectious SARS-CoV-2 were performed in the Galveston National Laboratory ABSL3 facility. Hamsters were intranasally infected with 105 pfu of WT or N679K SARS-CoV-2 in 100 μl. Infected hamsters were weighed and monitored for illness over 7 days. Hamsters were anesthetized with isoflurane and nasal washes were collected with 400 μl of PBS on endpoint days (2, 4, and 7 dpi). Hamsters were euthanized by CO2 for organ collection. Nasal wash and lung were collected to measure viral titer and RNA. Left lungs were collected for histopathology.
Hsdhan aura strain
Manufactured by Inotiv
The HsdHan:AURA strain is a laboratory animal model used in research. It serves as a genetic resource for studies. The core function of this strain is to provide a standardized animal model for scientific investigation.
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Lab products found in correlation
Antibiotic streptomycin, by Thermo Fisher Scientific (4 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Calu 3 cells, by American Type Culture Collection (2 mentions)
Ampicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Vero e6 cell, by American Type Culture Collection (1 mentions)
Penicillin streptomycin, by American Type Culture Collection (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
8 protocols using hsdhan aura strain
1
SARS-CoV-2 Infection Dynamics in Hamsters
2
Hamster and Primate Cell Culture
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The Syrian hamsters (HsdHan:AURA strain) were purchased from Envigo (Indianapolis, IN). African green monkey kidney epithelial Vero E6 cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) with 5% fetal bovine serum (FBS; HyClone Laboratories, South Logan, UT) and 1% antibiotic/ streptomycin (Gibco). Human lung adenocarcinoma epithelial Calu-3 2B4 cells were maintained in a high-glucose DMEM supplemented with 10% FBS and 1% penicillin/streptomycin at 37°C with 5% CO2. The EpiAirway system is a primary human airway 3D tissue model purchased from MatTek Life Science (Ashland, MA). This EpiAirway system was maintained with the provided culture medium at 37°C with 5% CO2 following the manufacturer’s instruction. All other culture medium and supplements were purchased from ThermoFisher Scientific (Waltham, MA). All cell lines were verified and tested negative for mycoplasma.
3
SARS-CoV-2 infection in Syrian hamsters
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Three-to four-week-old male Syrian hamsters (HsdHan:AURA strain) were purchased from Envigo. Animals were intranasally inoculated with 100 mL PBS (mock) or PBS containing 105 focus forming units (ffu) of WT or F17A mutant SARS-CoV-2. Animals were monitored for weight loss and signs of disease for 7 days. On 2 and 4 days post infection (dpi), cohorts of 5 animals were anesthetized with isoflurane and nasal washed with PBS and then sacrificed. Lung tissue was then taken to analyze viral titer by focus forming assay and pathology by hemoxylin and eosin (H&E) staining. H&E staining was performed by the University of Texas Medical Branch Histology Laboratory and then analyzed and scored by a blinded pathologist.
All animal studies were carried out in accordance with a protocol approved by the University of Texas Medical Branch Institutional Animal Care and Use Committee and complied with United States Department of Agriculture guidelines in a laboratory accredited by the Association for Assessment and Accreditation of Laboratory Animal Care. Procedures involving infectious SARS-CoV-2 were performed in the Galveston National Laboratory animal biosafety level 3 (ABSL3) facility.
All animal studies were carried out in accordance with a protocol approved by the University of Texas Medical Branch Institutional Animal Care and Use Committee and complied with United States Department of Agriculture guidelines in a laboratory accredited by the Association for Assessment and Accreditation of Laboratory Animal Care. Procedures involving infectious SARS-CoV-2 were performed in the Galveston National Laboratory animal biosafety level 3 (ABSL3) facility.
4
Hamster Vero E6 Cell Culture
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The Syrian golden hamsters (HsdHan:AURA strain) were purchased from Envigo (Indianapolis, IN). Vero E6 cells, an African green monkey kidney epithelial cell line (ATCC, Manassas, VA, USA), were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Gibco/Thermo Fisher, Waltham, MA, USA) with 10% fetal bovine serum (FBS; HyClone Laboratories, South Logan, UT) plus 1% ampicillin/streptomycin (Gibco). The authenticity of Vero E6 cells was verified using Short Tandem Repeat profiling by ATCC. The cells were tested negative for mycoplasma.
5
Characterizing SARS-CoV-2 in Human Airway Cells
6
Establishing SARS-CoV-2 Cell Culture Models
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The Syrian golden hamsters (HsdHan:AURA strain) were purchased from Envigo (Indianapolis, IN). African green monkey kidney epithelial Vero E6 cells (provided by R. Baric) were grown in Dulbecco’s modified Eagle’s medium (DMEM; Gibco/ThermoFisher, Waltham, MA, USA) with 10% fetal bovine serum (FBS; HyClone Laboratories, South Logan, UT) and 1% antibiotic/streptomycin (Gibco). Human lung adenocarcinoma epithelial Calu-3 cells (ATCC, Manassas, VA, USA) were maintained in a high-glucose DMEM containing sodium pyruvate and GlutaMAX (Gibco) with 10% FBS and 1% penicillin/streptomycin at 37°C with 5% CO2. The EpiAirway system is a primary human airway 3D tissue model purchased from MatTek Life Science. This EpiAirway system was maintained with the provided culture medium at 37 °C with 5% CO2 following the manufacturer’s instruction. All other culture medium and supplements were purchased from ThermoFisher Scientific (Waltham, MA). All cell lines were verified and tested negative for mycoplasma.
7
Hamster and Cell Lines for SARS-CoV-2 Research
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The Syrian hamsters (HsdHan:AURA strain) were purchased from Envigo (Indianapolis, IN). African green monkey kidney epithelial Vero E6 cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) with 5% fetal bovine serum (FBS; HyClone Laboratories, South Logan, UT) and 1% antibiotic/ streptomycin (Gibco). Human lung adenocarcinoma epithelial Calu-3 2B4 cells were maintained in a high-glucose DMEM supplemented with 10% FBS and 1 % penicillin/streptomycin at 37 °C with 5% CO2. The EpiAirway system is a primary human airway 3D tissue model purchased from MatTek Life Science (Ashland, MA). This EpiAirway system was maintained with the provided culture medium at 37 °C with 5% CO2 following the manufacturer’s instruction. All other culture medium and supplements were purchased from ThermoFisher Scientific (Waltham, MA). All cell lines were verified and tested negative for mycoplasma.
8
SARS-CoV-2 Infection in Golden Syrian Hamsters
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Three- to four-week-old male golden Syrian hamsters (HsdHan:AURA strain) were purchased from Envigo. All studies were conducted under a protocol approved by the UTMB Institutional Animal Care and Use Committee and complied with USDA guidelines in a laboratory accredited by the Association for Assessment and Accreditation of Laboratory Animal Care. Procedures involving infectious SARS-CoV-2 were performed in the Galveston National Laboratory ABSL3 facility. Hamsters were intranasally infected with 105 pfu of WT or NSP3 Mut1 or NSP3 Mut2 SARS-CoV-2 in 100 μl. Infected hamsters were weighed and monitored for illness over 7 days. Hamsters were anesthetized with isoflurane and nasal washes were collected with 400 μl of PBS on endpoint days (2, 4, and 7 dpi). Hamsters were euthanized by CO2 for organ collection. Nasal wash and lung were collected to measure viral titer and RNA. Left lungs were collected for histopathology.
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