CARD-CNB Cryopreservation Course Report

Organizers, instructors, lecturers and participants at the CARD-CNB Cryopreservation Course, held at CNB-CSIC in Madrid, Spain, on 7-11 October 2013 and organized by Naomi Nakagata (CARD-University of Kumamoto, Japan) and Lluis Montoliu (CNB-CSIC, Madrid, Spain)
Organizers, instructors, lecturers and participants at the CARD-CNB Cryopreservation Course, held at CNB-CSIC in Madrid, Spain, on 7-11 October 2013 and organized by Naomi Nakagata (CARD-University of Kumamoto, Japan) and Lluis Montoliu (CNB-CSIC, Madrid, Spain)

This past week, 7-11 October 2013, the CARD-CNB Mouse Sperm and Embryo Cryopreservation Course was held at CNB-CSIC, in Madrid, Spain, with great success and accompanied with also great sunny weather. This was the first cryopreservation course of this kind, co-organized by Naomi Nakagata (CARD-University of Kumamoto, Japan) and Lluis Montoliu (CNB-CSIC, Madrid, Spain), were the newest methods developed by CARD, at the Nakagata lab, were demonstrated in Europe, directly by the CARD team. The instructors at this CARD-CNB course were commanded by the CARD-University of Kumamoto team, from Japan (Toru Takeo, Kiyoko Fukumoto, Tomoko Kondo, Yukie Haruguchi, Yumi Takeshita, Yuko Nakamuta and Shuuji Tsuchiyama), and additional help and collaboration was provided from the Mouse Biology Program at UC-Davis, CA, USA (Kristy Kinchen), from INIA, Madrid, Spain (Raúl Fernández), from CIEMAT, Madrid, Spain (Jesús Martínez), from USA (Jorge Sztein), from Paratechs, Lexington, KY, USA (Barbara Stone) and from CNB-CSIC (Julia Fernández, María Jesús del Hierro, Marta Castrillo and Isabel Martín-Dorado), for several of the methods demonstrated. All instructors must to be praised for their deep knowledge, patience and extraordinary dedication and commitment towards the success of this course. Complementary and most interesting lectures were provided on a wide variety of topics related to the course main focus, including: animal welfare and regulations by Belén Pintado and Jorge Guillén; the history, fundaments and comparison of methods by Jorge Sztein; the effects of the in vitro culture of mouse embryos by Alfonso Gutiérrez-Adán; safety and handling issues of liquid nitrogen by Jesús Martínez; shipping frozen and refrigerated materials by Toru Takeo, databases in a cryopreservation lab, by Shuuji Tsuchiyama, about EMMA by Lluis Montoliu and CARD by Naomi Nakagata, as examples of mouse embryo and sperm archives, and, also, on the new editing nucleases for genome modification, by Kai Schönig (Mannheim, Germany), a talk sponsored by Sigma.

As many as 24 participants, coming from research institutions or companies located in 16 countries around the world (UK, Spain, Australia, USA, Canada, Czech Republic, Brazil, Finland, France, Denmark, Israel, Netherlands, Portugal, Sweden, Italy and Taiwan) were presented with the latest advances in mouse sperm and embryo cryopreservation and all associated mouse reproductive biology ancillary techniques.

The topics covered by the course included the following major areas: obtaining sperm from mouse cauda epididymis, obtaining unfertilized mouse oocytes, three different types of in vitro fertilization techniques (using fresh, frozen or refrigerated sperm), vitrification of unfertilized oocytes and 2-cell embryos (freezing and thawing), slow-method for freezing and thawing 2-cell embryos, refrigerated sperm and 2-cell embryos, abdominal and scrotal vasectomies, three types of embryo transfer (oviduct, uterus and non-surgical, with NSET tools), freezing and thawing mouse sperm and ICSI, among many additional common methodologies used to handle mouse embryos and gametes adequately.

The course was very intensive, but the kind atmosphere created by participants and instructors was excellent and, hence, all the tight and carefully devised demonstrations and practices, packed within a very busy schedule, could be run smoothly and successfully without problems. The vast experience in running this type of cryopreservation courses and the remarkable professionality of our colleagues from CARD-University of Kumamoto were key for the accomplished success. All methods followed their three-step learning process. At first, the theory and fundaments were briefly provided and summarized. Then, the method was demonstrated by instructors and, finally, the participants executed the procedures by themselves, with the help of instructors.

The participants left this cryopreservation course to return to their countries and institutions with the most satisfactory results obtained and with plenty of new information to digest, process and reproduce. All participants were given the task to spread the word and disseminate the use of these highly efficient and robust cryopreservation techniques that have boosted the field.

This CARDCNB cryopreservation course was sponsored by the International Society for Transgenic Technologies (ISTT) and received the co-sponsorship and support from a number of companies whose contributions need to be greatly acknowledged as well: Leica, Charles River, Sigma, Labotect, Cosmo-Bio, Kyudo, Harlan and Paratechs.

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