Protocols for Cryopreservation and Rederivation of Rat Gametes.
New genome-editing tools, such as ZFNs, TALEN, and CRISPR/Cas9, have enabled the generation of gene-modified models effectively in mammals. These technologies are a powerful tool for studying gene function and creating animal models for human diseases. On the other hand, such gene-modified animals are raised in numerous experimental animal facilities, which puts pressure on breeding space and maintenance costs. Embryo and sperm cryopreservation is not only the most simple and cost-effective method available for most gene-modified strains but also the most reliable method to preserve strains to avoid breeding problems and contamination. We have established a reliable, high quality embryo and sperm cryopreservation system for rat strains, ensuring the longevity of these valuable resources for the scientific community. These cryopreserved resources have been successfully used to rederive next generation pups using embryo transfer and intracytoplasmic sperm injection (ICSI). In this chapter, we describe in detail protocols for rat embryo vitrification and sperm cryopreservation followed by pup rederivation using the ICSI procedure and embryo transfer.
['Takizawa|Akiko|A|', 'Eto|Tomoo|T|']
D000818:Animals D015925:Cryopreservation D046149:Embryo Culture Techniques D004624:Embryo Transfer D004622:Embryo, Mammalian D047108:Embryonic Development D005260:Female D000072669:Gene Editing D008297:Male D051381:Rats D020554:Sperm Injections, Intracytoplasmic D013094:Spermatozoa
Cryopreservation Rederivation Sperm Embryo transfer ICSI Vitrification Two-cell