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Mouse Sperm Cryopreservation and Recovery using the I·Cryo Kit
Published on: December 12, 2011
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A simple and economic protocol for efficient in vitro fertilization using cryopreserved mouse sperm.
Magdalena Wigger1,2, Simon E Tröder1,2, Branko Zevnik1,2
1Cluster of Excellence Cellular Stress Responses in Aging-Associated Diseases (CECAD), Faculty of Medicine and University Hospital Cologne, University of Cologne, Cologne, Germany.
Plos One
|October 28, 2021
Summary
We developed a cost-effective method for in vitro fertilization using cryopreserved sperm from genetically engineered C57BL/6 mice. This approach simplifies rederivation and supports the 3Rs principles in mouse colony management.
Area of Science:
- Genetics and Genomics
- Animal Models
- Reproductive Biology
Background:
- Genome editing tools like CRISPR/Cas have increased genetically engineered mouse models.
- Sperm cryopreservation is preferred over embryo freezing for archiving and distributing mouse models.
- Current in vitro fertilization (IVF) protocols using cryopreserved sperm are efficient but costly due to specialized media.
Purpose of the Study:
- To develop a cost-effective and adaptable IVF protocol for cryopreserved C57BL/6 mouse sperm.
- To reduce the financial burden associated with routine IVF procedures for mouse rederivation.
- To provide a harmonized protocol supporting the 3Rs (Replacement, Reduction, Refinement) in animal research.
Main Methods:
- Modification of an affordable commercial fertilization medium.
- Detailed step-by-step protocol for reagent preparation and IVF procedure.
- Large-scale comparison of fertilization rates using the novel method versus current protocols.
Main Results:
- The novel SEcuRe (Simple Economical set-up for Rederivation) method demonstrated equal or superior fertilization rates compared to existing protocols.
- The protocol proved effective for genetically engineered C57BL/6 mouse lines.
- The approach significantly reduces costs associated with IVF media.
Conclusions:
- The SEcuRe method offers an affordable, adaptable, and efficient solution for rederivation using cryopreserved C57BL/6 sperm.
- This protocol facilitates broader application in mouse colony management, aligning with the 3Rs.
- The simplified approach reduces the complexity and cost of maintaining genetically engineered mouse models.
