Three groups of female NOD/ShiLtJ mice (stock 001976) and one group of female ICR/HaJ (stock 009122) mice were used (4 weeks; The Jackson Laboratory). Both NOD/ShiLtJ and ICR/HaJ mice are descended from ICR stock and carry the unique H2g7 MHC haplotype. However, ICR/HaJ mice do not develop diabetes and were used as controls. Blood glucose was measured weekly after 3-h fasting. At the beginning of the 14th week of age, three groups of NOD mice were treated with PBS, rAAV-GFP, and rAAV-mKL, respectively. The ICR/HaJ group was treated with PBS and served as controls. In brief, PBS, rAAV-GFP, or rAAV-mKL was carefully injected, at the dose of 2.57 × 109 viral genome copies per gram body wt (500 μL), into the region of pancreas starting from the splenic lobes toward the duodenal lobes of pancreas via intraperitoneal delivery (10 (link)). Glucose tolerance was tested at 7 and 14 days after gene delivery. Animals were killed at 16 days after gene delivery. Blood was collected for measuring plasma insulin levels. After perfusion with saline, pancreases were processed for fixation and paraffin embedding. The immunohistochemical staining for Klotho, insulin, and/or T-cell infiltration was performed as described below.
Nod shiltj mice
Manufactured by Jackson ImmunoResearch
Sourced in United States, Montenegro
The NOD/ShiLtJ mouse is a strain of laboratory mice that is commonly used in research. These mice have a genetic background that makes them susceptible to developing type 1 diabetes. They can be used as a model to study the underlying mechanisms of this autoimmune disease.
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Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Anti pd 1 antibody clone rmp1 14, by BioXCell (2 mentions)
Macrolide tylosin tartrate, by Merck Group (2 mentions)
Anti ctla4 clone 9d9, by BioXCell (1 mentions)
Migg2b, by BioXCell (1 mentions)
Diastix, by Bayer (1 mentions)
Nod scid, by Jackson ImmunoResearch (1 mentions)
Nod shiltj, by Jackson ImmunoResearch (1 mentions)
Balb c mice, by BioLASCO (1 mentions)
C57bl 6j b6 male mice, by Jackson ImmunoResearch (1 mentions)
Nor ltj, by Jackson ImmunoResearch (1 mentions)
Fetal bovine serum (fbs), by Greiner (1 mentions)
C57bl 6 mice, by Jackson ImmunoResearch (1 mentions)
Ot 2 mice, by Jackson ImmunoResearch (1 mentions)
C57bl 6, by Jackson ImmunoResearch (1 mentions)
Nod shijcl, by CLEA Japan (1 mentions)
Nod scid, by CLEA Japan (1 mentions)
Shuffle t7 competent e coli cells, by New England Biolabs (1 mentions)
B6.129s7 rag1tm1mom j mice, by Jackson ImmunoResearch (1 mentions)
El4 atcc tib 39 cells, by American Type Culture Collection (1 mentions)
50 protocols using nod shiltj mice
1
Klotho Gene Therapy for Diabetes in NOD Mice
2
NOD/ShiLtJ Mouse Model for Type 1 Diabetes
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Non-obese diabetic (NOD/ShiLtJ) mice, a polygenic model for T1D, were purchased from the Jackson Laboratory (Bar Harbor, ME, USA). NOD/ShiLtJ female mice exhibit diabetes beginning at around 16 weeks of age, characterized by insulitis, a leukocytic infiltrate of the pancreatic islets23 (link). Plasma glucose levels were used to determine disease onset. All animals for both studies were housed and bred in pathogen-free micro-isolator cages at the animal facilities operated by The Jackson Laboratory West (Sacramento, CA.). This study was conducted by In Vivo Services at The Jackson Laboratory Sacramento facility, an OLAW-assured and AAALAC-accredited organization. This study was performed according to the Jackson Laboratory IACUC-approved protocol and in compliance with the Guide for the Care and Use of Laboratory Animals (National Research Council, 1996 [Liposome study] and 2011 [HSA study]).
3
LPS-Induced Diabetes in NOD/ShiLTj Mice
4
Preventing Diabetes in NOD Mice
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Female nonobese diabetic (NOD)/ShiLtJ mice (The Jackson Laboratory, stock #001976) were maintained with free access to standard food and water. Nine-week-old female mice confirmed to be prediabetic (negative for glucosuria) were randomized and dosed with muC5H9v2 mAb, CX-630 Pb-Tx, or mouse IgG2a (BIW) by intraperitoneal injection. In the combination study, 5-week-old prediabetic female NOD mice were injected intraperitoneally with muC5H9v2 mAb, CX-630 Pb-Tx, or mIgG2a at 10 mg/kg on days 0, 4, and 7 in combination with either anti–CTLA-4 clone 9D9 (Bio X Cell, catalog #BP0164) or mIgG2b (Bio X Cell, catalog #BP0086) at 10 mg/kg. Body weights and urinary glucose levels were monitored daily. Urinary glucose was monitored daily using a glucose test strip (Diastix; Bayer Corporation). Blood obtained by means of a tail nick was tested for glucose with an Alpha TRAK 2 test strip inserted into a glucometer (Abbott Animal Care, now owned by Zoetis). Any animal with detectable glucose in urine (≥100 mg/dL) was immediately tested for blood glucose level. Animals with blood glucose levels ≥250 mg/dL for 2 consecutive days were designated as diabetic and euthanized by CO2 inhalation.
5
NOD/ShiLtJ Mouse Housing Protocol
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4 week-old female NOD/ShiLtJ mice were purchased from the Jackson Laboratory (Bar Harbor, ME) and housed 5 mice per cage in the Animal Care Services Biomedical Sciences Building SPF facility at the University of Florida, with food and water available ad libitum. All procedures were performed in accordance with guidelines and regulations put forth by the University of Florida Institutional Animal Care and Use Committee (UF IACUC) and according to a protocol approved by UF IACU.
6
Murine Diabetes Pathogenesis Study
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Female non-obese diabetic (NOD/ShiLtJ) mice were purchased from the Jackson Laboratory and were kept under specific pathogen-free conditions. All experimental protocols were approved by the Institutional Animal Care and Use Committee of the Forsyth Institute. All methods were carried out in accordance with the National Institutes of Health guidelines for the care and use of laboratory animals.
7
NOD Mouse Breeding and Maintenance
8
Mouse models for therapeutic evaluation
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Female C57BL/6 mice were purchased from the National Laboratory Animal Center (NLAC), NARLabs, Taiwan. All mice were maintained in the Laboratory Animal Center of Industrial Technology Research Institute (ITRI LAC) under specific pathogen-free (SPF) conditions. Healthy C57BL/6 mice at 6–12 weeks of age were used in experiments described in Figures 1 – 6
For the pharmacokinetics study, BALB/c mice were obtained from BioLASCO (Taiwan) and kept in the Development Center for Biotechnology, Taiwan. Healthy male mice at 6–8 weeks of age were used. For the biodistribution study, BALB/c mice were purchased from NLAC Labs. Mice were kept in the Institute of Nuclear Energy Research, Taiwan. Healthy male mice at 6–8 weeks of age were used.
Female NOD/ShiLtJ and NOD/ShiLtJNarl mice were obtained from The Jackson Laboratory (Bar Harbor, ME) and NLAC, respectively. Therapeutic experiments assessing the therapeutic efficacy of h145chIgGAA and h145CSA on NOD/ShiLtJ mice were conducted by The Jackson Laboratory-Sacramento Facility In Vivo Service, California. Therapeutic experiments employing NOD/ShiLtJNarl mice were performed in NLAC, Taiwan.
Mice were euthanized by inhalation of carbon dioxide in the euthanasia chamber.
For the pharmacokinetics study, BALB/c mice were obtained from BioLASCO (Taiwan) and kept in the Development Center for Biotechnology, Taiwan. Healthy male mice at 6–8 weeks of age were used. For the biodistribution study, BALB/c mice were purchased from NLAC Labs. Mice were kept in the Institute of Nuclear Energy Research, Taiwan. Healthy male mice at 6–8 weeks of age were used.
Female NOD/ShiLtJ and NOD/ShiLtJNarl mice were obtained from The Jackson Laboratory (Bar Harbor, ME) and NLAC, respectively. Therapeutic experiments assessing the therapeutic efficacy of h145chIgGAA and h145CSA on NOD/ShiLtJ mice were conducted by The Jackson Laboratory-Sacramento Facility In Vivo Service, California. Therapeutic experiments employing NOD/ShiLtJNarl mice were performed in NLAC, Taiwan.
Mice were euthanized by inhalation of carbon dioxide in the euthanasia chamber.
9
Generation and Analysis of Transgenic Mice
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C57BL/6J (B6) male mice, female non-obese diabetic (NOD/ShiLtJ) mice, female non-obese diabetes-resistant (NOR/LtJ), and INS1cre (B6(Cg)-Ins1tm1.1(cre)Thor/J) were obtained from the Jackson Laboratories. TMEM219flox/flox were generated in collaboration with Applied StemCell (Milpitas, CA) and housed at Charles River Laboratories (Wilmington, MA). TMEM219flox/floxINS1cre male mice (Beta-TMEM219−/−) were obtained by breeding TMEM219flox/flox mice with INS1cre mice, and the resulting colony was housed at Charles River Laboratories. Mice were housed at a temperature between 21 and 23 °C with 50–60% humidity and kept on a 12/12 h light/dark cycle with free access to food and water. All mice were cared for and used in accordance with institutional guidelines approved by the Boston Children’s Hospital Institutional Animal Care and Use Committee or in accordance with Italian law on animal care N° 116/1992 and the European Communities Council Directive EEC/609/86. All animal studies were approved by the Italian Ministry of Health and Local University of Milan Committee and by the Boston Children’s Hospital Institutional Animal Care and Use Committee. Some studies were conducted at the Jackson Laboratories.
10
Generation of Bone Marrow Dendritic Cells
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