breeding application for RIPDTR in NSG and BL6 background
Purpose: This application regards breeding of RIPDTR mouse line (in NSG or BL6 background strain) to create experimental and stock animals required for experimental applications like #25526: "Improving beta-cell function by drug treatment". NSG RIP-DTR mice lack mature T, B and NK cells and are suitable for studies where one does not want rejection / rejection of human cells and tissues in vivo. Also RIP-DTR mice express diphtheria toxin receptor (HBEGF).
Expected distress for the animals:
NSG;RIPDTR mice in this application are immunodeficient, thus do not have their own T, B or NK cells. Immunodeficient mouse strains are housed under SPF conditions and that all handling focuses on avoiding contact with human skin flora. Thus, expected distress in housing and breeding are therefore minimal, and not significantly different from housing and breeding of immunologically conventional mice. We do not foresee additional distress to these animals, except for what is occasionally associated with normal breeding. We will monitor all our breeding pairs to ensure animal well-being.
Expected scientific or societal benefit: The mouse strain is suitable for studies where one does not want rejection / rejection of human cells and tissues in vivo. Our projects aim at characterising the unknown mechanisms governing the decay of beta-cells in monogenic diabetes and to improve therapeutic strategies by increasing beta-cell resistance to cellular stress. This study will provide the first comprehensive cellular and molecular timeline of beta-cell decay as well as systematically define the beta-cell niche in diabetes. As only a limited number of scenarios can be considered for stress-related cell-fate changes, the revealed process of beta-cell failure will be relevant for the complex forms of diabetes. Moreover, our transcriptomic analyses will reveal new potential therapeutic targets. Our findings will allow a much more efficient clinical intervention with a greater gain in terms of functionality and health span for patients.
Number of animals to be used: a total of 1650 RIPDTR transgenic mice in NSG and BL6 background.
Replacement, reduction and improvement
We have good experience in breeding mouse strains that are immunodeficient. We therefore do not expect to increase breeding for other reasons than the use of the animals. By using our own breeding animals, we will be able to plan experiments more efficiently and reduce unnecessary purchases of animals.
All experiments that do not require a living organism (metabolism; systemic organ interactions) will be performed in vitro by using patient-derived hiPSC or isolated human islet cells. We will optimise the number of mice necessary for our projects by using common experimental groups, where the same set of collected samples will be processed separately for replying distinct questions, hence halving the number of required animals. Our breeding strategy takes into the consideration the reduction of unwanted transgenic genotypes by optimising the breeding pairs. Wherever possible, we will maintain the transgenic lines as homozygous, hence assuring the chance for mice with the desired genotype thus reducing the number of mice sacrificed due to unwanted genotype.
Expected distress for the animals:
NSG;RIPDTR mice in this application are immunodeficient, thus do not have their own T, B or NK cells. Immunodeficient mouse strains are housed under SPF conditions and that all handling focuses on avoiding contact with human skin flora. Thus, expected distress in housing and breeding are therefore minimal, and not significantly different from housing and breeding of immunologically conventional mice. We do not foresee additional distress to these animals, except for what is occasionally associated with normal breeding. We will monitor all our breeding pairs to ensure animal well-being.
Expected scientific or societal benefit: The mouse strain is suitable for studies where one does not want rejection / rejection of human cells and tissues in vivo. Our projects aim at characterising the unknown mechanisms governing the decay of beta-cells in monogenic diabetes and to improve therapeutic strategies by increasing beta-cell resistance to cellular stress. This study will provide the first comprehensive cellular and molecular timeline of beta-cell decay as well as systematically define the beta-cell niche in diabetes. As only a limited number of scenarios can be considered for stress-related cell-fate changes, the revealed process of beta-cell failure will be relevant for the complex forms of diabetes. Moreover, our transcriptomic analyses will reveal new potential therapeutic targets. Our findings will allow a much more efficient clinical intervention with a greater gain in terms of functionality and health span for patients.
Number of animals to be used: a total of 1650 RIPDTR transgenic mice in NSG and BL6 background.
Replacement, reduction and improvement
We have good experience in breeding mouse strains that are immunodeficient. We therefore do not expect to increase breeding for other reasons than the use of the animals. By using our own breeding animals, we will be able to plan experiments more efficiently and reduce unnecessary purchases of animals.
All experiments that do not require a living organism (metabolism; systemic organ interactions) will be performed in vitro by using patient-derived hiPSC or isolated human islet cells. We will optimise the number of mice necessary for our projects by using common experimental groups, where the same set of collected samples will be processed separately for replying distinct questions, hence halving the number of required animals. Our breeding strategy takes into the consideration the reduction of unwanted transgenic genotypes by optimising the breeding pairs. Wherever possible, we will maintain the transgenic lines as homozygous, hence assuring the chance for mice with the desired genotype thus reducing the number of mice sacrificed due to unwanted genotype.