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Open Access Comparison of Fertility Between Intracytoplasmic Sperm Injection and In Vitro Fertilization with a Partial Zona Pellucida Incision by Using a Piezo-Micromanipulator in Cryopreserved Inbred Mouse Spermatozoa

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Cryopreservation of mouse spermatozoa has been widely applied for maintenance of transgenic and knockout lines. However, the fertility of cryopreserved spermatozoa from some inbred strains such as C57BL/6 and BALB/c is extremely poor. We have recently reported that a partial zona-pellucida incision by piezo-micromanipulator (ZIP) significantly improves the fertilization rate and subsequent embryonic development after in vitro fertilization (IVF) using cryopreserved C57BL/6 transgenic mouse spermatozoa and that inbred C57BL/6 mice could be produced by intracytoplasmic sperm injection (ICSI). These findings prompted us to compare the efficiency of fertilization and subsequent embryonic development between ICSI and IVF with ZIP (ZIP/IVF) using cryopreserved spermatozoa. In conventional IVF, BALB/cA, C57BL/6J, and B6C3F1 cryopreserved spermatozoa fertilized 19%, 0%, and 51% of oocytes, respectively. The fertilization rates of manipulated oocytes by ICSI versus ZIP/IVF using cryopreserved BALB/cA spermatozoa were 52% versus 68%, cryopreserved C57BL/6J spermatozoa were 43% versus 63%, and cryopreserved B6C3F1 spermatozoa were 58% versus 82%, respectively. In these strains, fertilization rates for ZIP/IVF were significantly higher (P < 0.05) than for other techniques. However, embryonic development to term for oocytes fertilized by cryopreserved spermatozoa was not significantly different between ZIP/IVF and ICSI in C57BL/6J and B6C3F1. The overall efficiency of mouse production in ZIP/IVF was higher than for ICSI and conventional IVF in C57BL/6J and B6C3F1. Furthermore, ZIP/IVF required approximately 3.3 times less manipulation time than did ICSI. Our results indicate that ZIP is a useful assisted reproductive technique for IVF of ova by cryopreserved spermatozoa and improves production in some mouse strains.

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Document Type: Case Report

Affiliations: 1: Chugai Institute for Medical Science, Inc., Japan, Chugai Pharmaceutical Co., Ltd., Japan 2: Chugai Institute for Medical Science, Inc., Japan

Publication date: January 1, 2004

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  • The Journal of the American Association for Laboratory Animal Science (JAALAS) serves as an official communication vehicle for the American Association for Laboratory Animal Science (AALAS). The journal includes a section of refereed articles and a section of AALAS association news. The mission of the refereed section of the journal is to disseminate high-quality, peer-reviewed information on animal biology, technology, facility operations, management, and compliance as relevant to the AALAS membership. JAALAS accepts research reports (data-based) or scholarly reports (literature-based), with the caveat that all articles, including solicited manuscripts, must include appropriate references and must undergo peer review.

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