Inbred BALB/cJ and Nu/J 8-week-old female mice (Jackson Laboratory, Bar Harbor, USA) were housed under specific-pathogen-free conditions in individually ventilated cages (IVCs; Tecniplast, Milan, Italy) at the Animal Facility of the Unit of Biotechnology of Laboratory Animals. During infection, mice were assigned to groups of no more than six and housed in a biosafety level 2 (BSL2) ventilated rack with HEPA filters (IsoCage N; Tecniplast). All infection procedures were performed under a BSL2 laminar flow hood to comply with biosafety institutional rules. Animals received autoclaved fresh water and a standard autoclaved mouse diet ad libitum (Labdiet 5K67; PMI Nutrition, USA).
Isocage n
Manufactured by Tecniplast
Sourced in France, Germany
The IsoCage N is a laboratory equipment designed for housing laboratory animals. It provides a secure and controlled environment for the animals during research or experimentation.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using isocage n
1
Murine Infection Protocol for BSL2
2
BALB/c Mouse Model for Experimental Study
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A total of 56 BALB/cJ female mice (6–8 weeks old) were bred at the Transgenic and Experimental Animal Unit of Institut Pasteur de Montevideo under specific pathogen-free conditions in individually ventilated racks (IVC, 1285L, Tecniplast, Milan, Italy). During the experimental procedure, the mice were housed in groups of seven in negative pressure microisolators (ISOCageN, Tecniplast) with aspen chip wood bedding (Toplit 6, Safe, Augy, France), paper towels and cardboard tubes for environmental enrichment. They had ad libitum access to autoclaved food (5K67, LabDiet, MO, US) and filtered water. The housing environmental conditions during the experiment were as follows: 20 ± 1 °C temperature, 30–70% relative humidity, negative pressure (biocontainment) and a light/dark cycle of 14/10 h. All procedures were performed under Biosafety level II conditions. The mice were randomly distributed into three experimental groups: infected (n = 20), infected + IVM (n = 20) and control (n = 16) groups. Experiments were conducted in three independent replicates.
3
C57BL/6 Mouse Housing and Acclimation
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Female C57BL/6JOlaHsd (C57BL/6) mice were purchased from Envigo RMS GmbH, Venray, Netherlands. Mice were housed under pathogen-free conditions in individually ventilated cages type Sealsafe Plus GM500 or IsoCage N Biocontainment system (Tecniplast, Hohenpeißenberg, Germany). Sterilized water and food were provided ad libitum. The experiments were started after 7 days of acclimatization of the mice. Treatment of the mice was always done under isoflurane anesthesia.
4
Humanized Mouse Models for SARS-CoV-2 Infection
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hACE2-KI mice (B6.Cg-Ace2tm1(ACE2)Dwnt) and hACE2-K18Tg mice (Tg(K18-hACE2)2Prlmn) were described previously3 (link),18 (link). All mice were produced at the specific-pathogen-free facility of the Institute of Virology and Immunology (Mittelhäusern), where they were maintained in individually ventilated cages (blue line, Tecniplast), with 12-h:12-h light:dark cycle, 22 ± 1 °C ambient temperature and 50 ± 5% humidity, autoclaved food and acidified water. At least 7 days before infection, mice were placed in individually HEPA-filtered cages (IsoCage N, Tecniplast). Mice (10 to 12 weeks old) were anaesthetized with isoflurane and infected intranasally with 20 μl per nostril with the virus inoculum described in the results section. One day after inoculation, infected hACE2-K18Tg mice were placed in the cage of another hACE2-K18Tg contact mouse. Mice were monitored daily for bodyweight loss and clinical signs. Oropharyngeal swabs were collected under brief isoflurane anaesthesia using ultrafine sterile flock swabs (Hydraflock, Puritan, 25-3318-H). The tips of the swabs were placed in 0.5 ml of RA1 lysis buffer (Macherey-Nagel, 740961) supplemented with 1% β-mercaptoethanol and vortexed. At 2 or 4 dpi, mice were euthanized, and organs were aseptically dissected. Systematic tissue sampling was performed as detailed previously3 (link).
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