title: Aseptic minimum volume vitrification technique for porcine parthenogenetically activated blastocyst.
abstract: Minimum volume vitrification may provide extremely high cooling and warming rates if the sample and the surrounding medium contacts directly with the respective liquid nitrogen and warming medium. However, this direct contact may result in microbial contamination. In this work, an earlier aseptic technique was applied for minimum volume vitrification. After equilibration, samples were loaded on a plastic film, immersed rapidly into factory derived, filter-sterilized liquid nitrogen, and sealed into sterile, pre-cooled straws. At warming, the straw was cut, the filmstrip was immersed into a 39 degree C warming medium, and the sample was stepwise rehydrated. Cryosurvival rates of porcine blastocysts produced by parthenogenetical activation did not differ from control, vitrified blastocysts with Cryotop. This approach can be used for minimum volume vitrification methods and may be suitable to overcome the biological dangers and legal restrictions that hamper the application of open vitrification techniques.
authors.affiliation: BGI/HuaDa Shenzhen, Shenzhen, China. linlin@genomics.org.cn
authors.forename: Lin
authors.initials: L
authors.lastname: Lin
authors.forename: Yutao
authors.initials: Y
authors.lastname: Yu
authors.forename: Xiuqing
authors.initials: X
authors.lastname: Zhang
authors.forename: Huanming
authors.initials: H
authors.lastname: Yang
authors.forename: Lars
authors.initials: L
authors.lastname: Bolund
authors.forename: Henrik
authors.initials: H
authors.lastname: Callesen
authors.forename: Gábor
authors.initials: G
authors.lastname: Vajta
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology*; D003080:Cold Temperature; D015925:Cryopreservation; D003451:Cryoprotective Agents; D003470:Culture Media; D005260:Female; D066298:In Vitro Techniques; D009584:Nitrogen; D009865:Oocytes / Q000166:cytology; D010312:Parthenogenesis; D013242:Sterilization; D013552:Swine; D058989:Vitrification
chemical_list: D003451:Cryoprotective Agents; D003470:Culture Media; D009584:Nitrogen