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5 protocols using 1 mm cat catheter

1

Ethanol-Induced Colitis Model in NSG Mice

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NSG mice were obtained from Charles River Laboratories [Sulzfeld, Germany]. Mice were kept under specific pathogen-free conditions in individually ventilated cages in a facility controlled according to the Federation of Laboratory Animal Science Association [FELASA] guidelines. Following engraftment on Day 1, mice were presensitised by rectal application of 150 µL of 10% ethanol on Day 8 using a 1-mm cat catheter [Henry Schein, Hamburg, Deutschland]. The catheter was lubricated with Xylocain©Gel 2% [AstraZeneca, Wedel]. Rectal application was performed under general anaesthesia using 4% isoflurane. Post-application mice were kept at an angle of 30° to avoid ethanol dripping. On Day 15, the mice were challenged by rectal application of 50% ethanol following the protocol used on Day 8. For the drug treatments, DES1 [D. E. Shaw Research, New York, USA] and tofacitinib [Merck KGaA, Darmstadt, Germany] were administered daily over one period of 3 days and a second period of 4 days [Days 7–9 and 14–17, respectively], such that each period encompassed one of the two ethanol challenges [on Days 8 and 15]. The dosages were 5 mg/kg/day for DES1 and 5 mg/kg/day for tofacitinib. The dosage of infliximab [Janssen Biotech, Horsham, Pennsylvania, USA] was 6 mg/kg/day, and treatment was limited to Days 7 and 14 only. All therapeutics were dissolved in PBS and administered intraperitoneally.
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2

Ethanol-induced Colitis Model in NOD-scid IL-2Rγ Mice

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NOD.cg-PrkdcSCID Il2rgtm1Wjl/Szj mice (abbreviated as NOD-scid IL-2Rγnull) were obtained from Charles River Laboratories (Sulzfeld, Germany). Mice were kept under specific pathogen-free conditions in individually ventilated cages. The facility was controlled according to the Federation of Laboratory Animal Science Association (FELASA) guidelines. Following engraftment (day 1), mice were pre-sensitized through rectal application of 150 µl of 10% ethanol on day 8 using a 1-mm cat catheter (Henry Schein, Hamburg, Germany). The catheter was lubricated with Xylocain© Gel 2% (AstraZeneca, Wedel). Rectal application was performed under general anesthesia using 4% isoflurane. Post application, mice were kept at an angle of 30° to avoid ethanol dripping. On days 15 and 18, mice were challenged with rectal application of 50% ethanol following the protocol described for day 8. Mice were killed on day 21. Pitrakinra (10 µg in 0.5% methylcellulose, 0.05% Tween-80) in PBS (Zadeh-Khorasani et al., 2013 (link)) was applied on days 7-9 and 14-21. In these groups, sterile saline (B. Braun Melsungen AG, Germany) served as control. Infliximab [6 mg/kg (Remicade©, Janssen, The Netherlands)] was applied on days 7, 14 and 17. An isotype antibody (human IgG1, kindly provided by, MorphoSys AG) was used as control. All treatments were applied intraperitoneally.
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3

Ethanol-Induced Colitis Model in NOD IL-2Rγnull Mice

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NOD IL-2Rγnull mice were obtained from Charles River Laboratories (Sulzfeld, Germany). Mice were kept under specific pathogen-free conditions in individually ventilated cages in a facility controlled according to the Federation of Laboratory Animal Science Association (FELASA) guidelines. Following engraftment (day 1) mice were pre-sensitized by rectal application of 150 µl of 10% ethanol on day 8 using a 1 mm cat catheter (Henry Schein, Hamburg, Germany). The catheter was lubricated with Xylocaine©Gel 2% (AstraZeneca, Wedel). The rectal application was performed under general anesthesia using 4% isoflurane. Post application mice were kept at an angle of 30° to avoid ethanol dripping. On day 15 and 18 mice were challenged by rectal application of 50% ethanol following the protocol of day 8. Mice were sacrificed on day 21. Pitrakinra (10 µg in 0.5% Methylcellulose, 0.05% TWEEN 80 in PBS) [34 (link)] was applied on day 7–9 and 14–21. Sterile Saline (B. Braun Melsungen AG, Germany) served as a control. Infliximab, [6 mg/kg (Remicade©, Janssen The Netherlands)] and isotype control (30 µg in 200 µl PBS, Morphosys AG, Planegg, Germany) were applied on day 7 and 14. All treatments were applied intraperitoneally.
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4

Ethanol-Induced Colitis Model in NSG Mice

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NSG mice were obtained from Charles River Laboratories (Sulzfeld, Germany). Mice were kept under specific pathogen–free conditions in individually ventilated cages in a facility controlled according to Federation of Laboratory Animal Science Association (FELASA) guidelines. After engraftment on day 1, mice were presensitized by rectal application of 150 µL of 10% ethanol on day 8 using a 1-mm cat catheter (Henry Schein, Hamburg, Germany). The catheter was lubricated with Xylocain Gel 2% (AstraZeneca, Wedel, Germany). Rectal application was performed under general anesthesia using 4% isofluran. Postapplication, mice were kept at an angle of 30° to avoid ethanol dripping. On day 15, mice were challenged by rectal application of 50% ethanol following the protocol of day 8. On day 18, mice were killed. Adalimumab was provided by Sanofi-Aventis Deutschland GmbH (Frankfurt am Main, Germany) and was applied in PBS (30 mg/kg/d) on days 7 and 14. The control groups were injected with 30 mg/kg of the isotype antibody (Sanofi-Aventis Deutschland GmbH, Frankfurt am Main, Germany) on days 7 and 14.
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5

Ethanol-Induced Colitis Model in NOD/scid IL-2Rγ Mice

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NOD/scid IL-2Rγnull mice were obtained from Charles River Laboratories (Sulzfeld, Germany). Mice were kept under specific pathogen-free conditions in individually ventilated cages in a facility controlled according to the Federation of Laboratory Animal Science Association guidelines. Following engraftment (day 1), mice were presensitized by rectal application of 150 µL of 10% ethanol on day 8 using a 1-mm cat catheter (Henry Schein, Hamburg, Germany). The catheter was lubricated with Xylocain Gel 2% (AstraZeneca, Wedel). Rectal application was performed under general anaesthesia using 4% Isofluran. Postapplication, mice were kept at an angle of 30° to avoid ethanol dripping. On days 15 and 18, mice were challenged by rectal application of 50% ethanol following the protocol of day 8. Mice were killed on day 21.
Mice were treated by intraperitoneal application of 30 µg of anti-CD1a antibody (Clone OKT6, BioXcell, West Lebanon, USA) or 30 µg of isotype control (Biolegend, San Diego, CA, USA) on days 7, 14, and 17.
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