A total of 100 plaque forming unit (PFU) of MHV-3 was injected intraperitoneally into the mice to establish a FVH model. MHV-3 was obtained from the American Type Culture Collection (Manassas, VA) and expanded according to a previously published protocol.14 (link) APAP (500 mg/kg) was used to establish a mouse model of non–virus-caused ALF. NETs were depleted by intraperitoneal injection of DNase I (750 U), a widely used method to remove the DNA scaffold of NETs,8 (link) at 4 hours and 48 hours after MHV-3 infection. For neutrophil depletion, mice were injected intraperitoneally with anti-Ly6G (clone 1A8; BioXcell, West Lebanon, NH) and rat IgG2a (clone 2A3; BioXcell, West Lebanon, NH) were used as an isotype control. To adoptively transfer neutrophils, BM neutrophils were purified using the Neutrophil Isolation Kit (Miltenyi Biotec, San Diego, CA). A total of 4 × 106 WT or fgl2-/- neutrophils were adoptively transferred into MHV-3–infected fgl2-/- mice by intravenous injection.
Mhv 3
Manufactured by American Type Culture Collection
Sourced in United States
The MHV-3 is a laboratory equipment designed for the cultivation and propagation of murine hepatitis virus (MHV) in cell culture. It provides a controlled environment for the growth and maintenance of this virus, which is commonly used in research applications.
Automatically generated - may contain errors
4 protocols using mhv 3
1
Establishing MHV-3 and APAP-Induced Liver Failure Models in Mice
2
Murine Hepatitis Virus Infection Model
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MHV-3 was obtained from the American Type Culture Collection (Manassas, VA, USA) and was plaque-purified on a monolayer of delayed brain tumor cells and titer-tested on L2 cells according to a standard plaque assay. Mice were injected intraperitoneally with MHV-3 (100 PFU/mouse) in saline solution (200 μl). For alanine transaminase and aspartate transaminase measurements, blood was collected to isolate serum at the indicated time after virus infection. Ten to twenty mice were used for the infection experiments.
3
Murine Hepatitis Virus-Induced Fulminant Hepatic Failure
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MHV-3 was obtained from the American Type Culture Collection (Manassas, VA), plaque purified on monolayer of delayed brain tumor cells (DBT), and titer tested on L2 cells according to a standard plaque assay [14 (link), 15 (link)]. DBT cells and L2 were provided generously by Dr. Gary A Levy (Toronto General Hospital, Toronto). Female BALB/cJ mice at 6–8 weeks old, weighed 18-20 g, were purchased from HFK Bioscience (Beijing). Viral fulminant hepatic failure was established by intraperitoneal injection of either 20 or 100PFU of MHV-3. In detail, MHV-3 was diluted in sterile PBS at a concentration of 100PFU/ml or 500 PFU/ml. Mice were injected intraperitoneally with MHV-3 (20 PFU or 100 PFU per mouse) in a total volume of 200 μl. Mice were anaesthetized by exposure to 0.08 ml/l aether (1.9%) and sacrificed by cervical dislocation for sample collection at 48 h post viral injection. Animal care and procedures on animal were in compliance with the guidelines outlined in the Guide for the Care and Use of Laboratory Animals and approved by the Committees on Animal Experimentation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology.
4
Murine Fulminant Hepatitis Model via MHV-3 Induction
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Female BALB/c mice (HFK Bioscience Company Ltd., Beijing, China) at 6-8 weeks of age were used in all animal experiments. The mice were housed in a controlled environment (specific pathogen-free, 12 h light/dark cycle, 21 ± 2 °C, humidity 50 ± 10%) and had free access to food and water. Fgl2-/- mice were generated by introducing a deletion at the N-terminal of the open reading frame of the Fgl2 locus using CRISPR/Cas9 technology. WT BALB/c mice were used as controls. To establish the fulminant hepatitis model, 100 plaque-forming unit (PFUs) of MHV-3 (American Type Culture Collection, Manassas, VA, USA) dissolved in 200 µL saline was injected into the WT and Fgl2-/- mice intraperitoneally. To deplete macrophages, mice were intravenously injected with 200 µL clodronate liposomes or phosphate-buffered saline (PBS)-liposomes as a control (Liposome B.V., the Netherlands) 24 h prior to MHV-3 injection. To inhibit MoMF infiltration, a chemokine receptor-2 (CCR2) inhibitor (cenicriviroc; 10 mg/kg) was intraperitoneally injected 24 h prior to MHV-3 infection. WT or Fgl2-/- mice were randomly selected to be sacrificed at 0, 24, 48, and 72 h following MHV-3 injection, and liver and blood samples were harvested for analysis.
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