Generation of rats from vitrified oocytes with surrounding cumulus cells via in vitro fertilization with cryopreserved sperm. (2nd August 2016)

• Record Type: Journal Article
• Authors: Fujiwara, Katsuyoshi; Kamoshita, Maki; Kato, Tsubasa; Ito, Junya; Kashiwazaki, Naomi
• Is Part Of: Animal science journal. Volume 88:Number 1(2017:Jan.)
• Page Start: 180
• Page End: 184
• Publication Date: 2016-08-02
• Subjects: in vitro fertilization -- oocyte vitrification -- rat -- sperm cryopreservation; Animal culture -- Periodicals; Livestock -- Periodicals; 636
• DOI: 10.1111/asj.12666
• Languages: English
• ISSNs: 1344-3941

Abstract

Abstract: The objective of this study was to evaluate fertility and full‐term development of rat vitrified oocytes after in vitro fertilization (IVF) with cryopreserved sperm. Oocytes with or without surrounding cumulus cells were vitrified with 30% ethylene glycol + 0.5 mol/L sucrose + 20% fetal calf serum by using the Cryotop method. The warmed oocytes were co‐cultured with sperm. Although the denuded/vitrified oocytes were not fertilized, some of the oocytes vitrified with cumulus cells were fertilized (32.7%) after IVF with fresh sperm. When IVF was performed with cryopreserved sperm, vitrified or fresh oocytes with cumulus cells were fertilized (62.9% or 41.1%, respectively). In addition, to confirm the full‐term development of the vitrified oocytes with surrounding cumulus cells after IVF with cryopreserved sperm, 108 vitrified oocytes with two pronuclei (2PN) were transferred into eight pseudopregnant females, and eight pups were obtained from three recipients. The present work demonstrates that vitrified rat oocytes surrounded by cumulus cells can be fertilized in vitro with cryopreserved sperm, and that 2PN embryos derived from cryopreserved gametes can develop to term. To our knowledge, this is the first report of successful generation of rat offspring derived from vitrified oocytes that were fertilized in vitro with cryopreserved sperm.