CARD – IP Mouse Sperm and Embryo Cryopreservation Course, Institut Pasteur, Paris, France, 20-24 June 2016

 

The International Society for Transgenic Technologies (ISTT) will proudly co-sponsor the CARD – IP Mouse Sperm and Embryo Cryopreservation Course that will be held at the renowned Pasteur Institute, in Paris, on 20-24 June 2016, organized by Naomi Nakagata (CARD-Kumamoto University, Japan, Coordinator of CARD) and Jean Jaubert (Pasteur Institute, Paris).

Recent developments from the laboratory of Prof. Naomi Nakagata (CARD-Kumamoto University, Japan) have pushed the envelope of mouse cryopreservation: i) improved female superovulation method; ii) fresh, frozen and cold storage sperm techniques; iii) optimized IVF methods. These improvements (see main references in program) have resulted in an unparalleled increase in efficiency of cryopreservation and rescue of relevant mouse lines.

The aim of this course is to introduce the newest CARD methods to researchers and technicians involved in mouse archiving and/or managing transgenic facilities and who are willing to implement these new methods in their work. These techniques will be taught directly by the team that devised them.

This course is open to anyone interested. Pre-application will be required, including, at least, a recent CV and a letter prepared by the intended participant describing how the applicant will benefit by attending this course and how relevant is the course material to his/her work. Additional documents are welcome, at the discretion of participants, including supporting letters by supervisors (where appropriate), reference letters, etc… A copy of the passport is mandatory. Applications should be submitted online, and will close on March 25th 2016.

The maximum number of participants attending this course will be 20, distributed among countries and institutions, and according the documentation provided and the interests expressed. Review and selection of participants will be done by the Teaching Committee and results will be communicated by April 15, 2016. The official language of the course will be English.

In addition to practical sessions, the course will also include several lectures of related interesting topics for the participants delivered by experts in each field.

See more at: http://www.pasteur.fr/fr/enseignement/ateliers/mouse-sperm-and-embryo-cryopreservation-course#sthash.BKcGh9VV.dpuf

 

Screen Shot 2016-02-02 at 15.20.55

Hands-on topics:

 

Making pipettes and embryo handling

Superovulating/Ultra-superovulating female mice

Isolating unfertilized mouse oocytes

Isolating and cold storage/shipping of mouse cauda epididymis

Freezing/thawing mouse sperm and IVF

Fresh mouse sperm and IVF

Freezing/thawing 2-cell IVF-derived mouse embryos

Vitrification of mouse oocytes and embryos

Embryo transfer techniques in mice (oviduct, uterus, NSET)

Vasectomy of male mice (scrotal and abdominal)

 

Additional lectures:

 

The laboratory mouse origin

Historic and scientific perspectives of transgenesis methods and the future of transgenic platforms

Historic and Scientific perspectives of embryo and sperm cryopreservation

Comparing current embryo and sperm cryopreservation methods

Vitrification of oocytes and their use for IVF

Cold storage and transport of germplasm

Shipping mice, refrigerated and frozen material

Managing and handling information in cryopreservation centers

CRISPR/Cas9 and the challenges in freezing these new GEM’s

NSET: non-surgical embryo transfer

Breeding, genotyping and back-ups of GEM’s

CARD

 

Instructors:

 

Naomi Nakagata (CARD-Kumamoto University, Japan)

Toru Takeo (CARD-Kumamoto University, Japan)

Shuuji Tsuchiyama (CARD-Kumamoto University, Japan)

Kiyoko Fukumoto (CARD-Kumamoto University, Japan)

Yukie Haruguchi (CARD-Kumamoto University, Japan)

Tomoko Kondo (CARD-Kumamoto University, Japan)

Yumi Takeshita (CARD-Kumamoto University, Japan)

Yuko Nakamuta (CARD-Kumamoto University, Japan)

Tomoko Umeno (CARD-Kumamoto University, Japan)

Hidetaka Yoshimoto (CARD-Kumamoto University, Japan)

Ayumi Mukunoki (CARD-Kumamoto University, Japan)

Mari Iwamoto (CARD-Kumamoto University, Japan)

Fumi Takahashi (CARD-Kumamoto University, Japan)

Kristy Kinchen (Gainesville, FL, USA)

Jean Jaubert (Institut Pasteur, France)

Franck Bourgade (Institut Pasteur, France)

Angélique Vincent (Institut Pasteur, France)

Claire Lecestre (Institut Pasteur, France)

Jorge Sztein (Barcelona, Spain)

Lluís Montoliu (CNB-CSIC, Madrid, Spain)

Barbara Stone (ParaTechs, Lexington KY, USA)

 

Additional lectures:

 

Naomi Nakagata (CARD-Kumamoto University, Japan)

Toru Takeo (CARD-Kumamoto University, Japan)

Shuuji Tsuchiyama (CARD-Kumamoto University, Japan)

Jorge Sztein (Barcelona, Spain)

Lluís Montoliu (CNB-CSIC, Madrid, Spain)

Fernando Benavides (MD Anderson, Smithville, USA)

Francina Langa Vives (Institut Pasteur, Paris, France)

Michel Cohen-Tannoudji (Institut Pasteur, Paris, France)

Xavier Montagutelli (Institut Pasteur, Paris, France)

Jean Jaubert (Institut Pasteur, Paris, France)

Barbara Stone (ParaTechs, Lexington KY, USA)

 

For any further information contact: enseignement@pasteur.fr

 

Tags: Cryopreservation course, embryo cryopreservation, ISTT co-sponsorisation, IVF, practical course, sperm cryopreservation, superovulation, vasectomy, vitrification

Report on the 2nd Oceania Transgenic Technology/Cryopreservation Symposium

2nd oceania symposium

The 2nd Oceania Transgenic Technology and Cryopreservation Symposium was held at the University of Tasmania’s Medical Sciences Precinct, Hobart, Australia on the 18th-19th of November, 2015. The meeting was a great success with for 48 participants from 23 research institutions across Australia, New Zealand, Japan and the USA and with the support of 9 sponsoring companies. The Organising Committee comprised of: Paul Scowen-chair and host (UTAS), Elizabeth Williams (University of Queensland Biological Resources), Kevin Taylor (Australian BioResources,) Irma Villaflor (Children’s Medical Research Institute, Westmead), Tanya Templeton (Australian Phenomics Network, Monash) and Karen Brennan (Victor Chang Cardiac Research Institute). Once again, it provided an opportunity for networking, for keeping up to date with the latest developments in transgenic technologies and for sharing knowledge through expertise, round table discussions and hands-on experience.

Highlights of the Programme:

The symposium began with a warm welcome from the host, Paul Scowen, and quickly moved on, beginning with a session of talks related to assisted reproduction techniques and their applications. Dr. Toru Takeo (Kumamoto University, Japan) , started by showing that ‘ultra-ovulation’ of female mice through the administration of a novel anti-serum (soon available as a commercial product) allows for consistent collection of a large number of usable oocytes from a single mouse. Combined with the IVF media products developed by the same group, mouse IVF techniques are now even further enhanced. Elizabeth Williams (UQBR) demonstrated that the CARD embryo vitrification technique can be adapted to use either straws or vials as the cryostorage vessel, allowing facilities to update their freezing techniques without having to change their liquid nitrogen storage equipment. An entertaining presentation was delivered by Prof. John McLaughlin, showing that cryovials can be easily converted with the new ‘Cryofork spatula’ to allow for easy vitrification of embryos in small media volumes. Rodrick Rupan described the challenges faced in rederiving immunodeficient colonies and how they were overcome. Continuing the rederivation and IVF theme, Mary-anne Migotto explained how UQBR have used IVF techniques to offer a rapid and large scale rederivation service. Tanya Templeton expanded upon her talk from the first Oceania symposium, giving an update on developments in ICSI technique and use of piezo methodology at Monash University. The session concluded with Julie Stanley’s talk on sperm cryopreservation at WEHI.

 

The second session covered various aspects of rodent health screening, biosecurity, monitoring, risk assessment and challenges when dealing with genetically modified mice models. Dr Trasti presented key concepts for a rodent sentinel program, establishing exclusion criteria, sample collection, interpretation of report and preparation of an action plan in case of an outbreak situation. Dr Villaflor discussed recommendations on which pathogens to monitor, issues with working with humanized immune deficient mice, quality control system and her experiences with adapting the latest trend in laboratory animal health monitoring programs. Dr Stevenson shared his insights on an appropriate scoring system for determining what agents and factors pose a risk to animal facilities housing genetically modified animals.

 

The third and final session of the day covered aspects of quality control. Sue Raboczyj (UQBR) and John Swift (OHIO) explained clearly the Quality Assurance and Control requirements for operating transgenic facilities with multiple clients, and ensuring the ongoing maintenance of a growing archive of cryopreserved material. Dr. Takeo also presented the methods used to establish a robust infrastructure at CARD (Kumamoto University). The day concluded with a personal account of undergoing the NATA accreditation process from Barbara Hunt (ANU).

Day 2 began with an extremely informative session on CRISPR technology. Kevin Taylor began with a talk on the introduction to CRISPR, the theory behind its evolution, the applications and advances based on his experiences at ABR in Moss Vale. Kevin also mentioned about the new refinements such as SCR7 and also briefly talked about electroporation. Fabien Delerue (UNSW) reported on the traditional methods utilised for creating knock-outs versus CRISPR/Cas9 and followed on with details of some of the projects that he is undertaking at UNSW and what he has done to refine how he carries out his projects being that he is a small outfit. Sandie Piltz’s work at the University of Adelaide is one of the pioneering Australian groups to start working with this technology, she discussed her personal experience with this methodology and what they have done to refine the technique, such as donor strains and ages, recipient strains, needle parameters, injection reagents and concentrations, environmental influences and superovulation techniques. Fiona Waters (WEHI) gave an introduction on the CRISPR/Cas9 methodology and then launched into the experiences from the WEHI team on how they have utilised and transferred their skills to this technique and the refinements they have made. Dirk Truman (APN) talked about the services that the APN provides, their efficiencies and that it is one of the first non-for-profit services offering this type of genome editing to Australian Researchers. Dirk also discussed the different species that are being used for this type of genome editing, off target effects, and what APN has done to refine the technique. The last talk for this session, Michelle Brownlee (ABR), talked about the day in the life of a microinjectionist. She discussed the strains of mice used, superovulation techniques, injection techniques, embryo transfer methods, refinements and troubleshooting.

The following session encompassed the administration and training aspects of working in a transgenic animal facility. Tracy Doan (UQBR) gave an overview of the database systems used and administrative support provided to help coordinate the cryopreservation and rederivation services at the Transgenic Animal Service of Queensland. Kevin Taylor (ABR) and Keri Smith (UTas) presented their respective training programs based on structure which also opened up lively discussions on different training opportunities offered in various institutions and suggestions to meet training needs of staff members. Dr David Steele who is Institutional Biosafety Committee (IBC) Chairman at the University of Tasmania, outlined the structure and important role and function of the IBC, including dealing with GMOs when conducting research in accordance with legislation, codes of practice and licensing requirements.

It is envisioned that future meetings will serve to strengthen our collaboration efforts with colleagues from various institutions doing the same type of work.

TT2016 Deadlines and Poster

TT2016, to be held in Prague in late March, should prove to be an exciting meeting. The scientific portion of the meeting will be from 20-23 March. In addition, there are three hands-on workshops that are available, including a Mouse Cryopreservation Workshop (16-18 March), a course on CRISPR/Cas9 programmable nucleases (16-18 March) and a Fish Transgenesis Course (23-25 March).

Early registration will close on 15 December, 2015, so register soon! Please note that ISTT members have discounted registrations, and can apply for a limited number of registration and travel awards (as detailed in e-mails sent to ISTT members). Applications and documentation for these awards must be sent in by 15 December, 2015 to istt@transtechsociety.org.

We also encourage you to submit an abstract for the poster session. A number of poster abstracts will be chosen for oral presentation, so this is an excellent opportunity to share your science. Poster submissions are also eligible for the TT2016 Poster awards. Abstracts are due by 15 December, 2015

Please note that a printable poster has been appended to this blog post. Feel free to print and post widely! Looking forward to seeing you at TT2016!

TT2016_poster A3w workshops

Ultra-superovulation in C57BL/6 mice: 100 oocytes/female

New achievement in Reproductive Biology by Toru Takeo & Naomi Nakagata (CARD, University of Kumamoto, Japan). Ultra-superovulation of C57BL/6 mice: 100 oocytes/female obtained priming the animals with equine chorionic gonadotropin (eCG) and inhibin antiserum (IAS)
New achievement in Reproductive Biology by Toru Takeo & Naomi Nakagata (CARD, University of Kumamoto, Japan). Ultra-superovulation of C57BL/6 mice: 100 oocytes/female obtained priming the animals with equine chorionic gonadotropin (eCG) and inhibin antiserum (IAS)

Last September, in Buffalo (USA), at the CARD-RPCI Mouse Sperm and Embryo Cryopreservation course organized by Naomi Nakagata, Aimee Stablewski and Jan Parker-Thornburg, Naomi Nakagata himself presented the preliminary results of an amazing achievement in Reproductive Biology they had accomplished at the University of Kumamoto (Japan), namely: the obtention of more than 100 oocytes per C57BL/6 female after devising a new protocol for superovulation, a method they introduced as ULTRA-superovulation. Now, these totally unexpected results see the light in the form of a scientific manuscript, published yesterday in the PLOS ONE journal:

Toru Takeo & Naomi Nakagata (2015) Superovulation Using the Combined Administration of Inhibin Antiserum and Equine Chorionic Gonadotropin Increases the Number of Ovulated Oocytes in C57BL/6 Female Mice. PLOS ONE, Published: May 29, 2015DOI: 10.1371/journal.pone.0128330

In brief, in this publication, Toru Takeo and Naomi Nakagata describe their superovulation results using young (4-weeks old) C57BL/6 female after envisaging a new priming protocol. The combined used of equine chorionic gonadotropin (eCG) and inhibin antiserum (IAS), in a protocol they call IASe treatment, significantly increased the number of oocytes obtained per C57BL/6 females. On average, more than 100 oocytes/female were obtained, about 3-4 times the number of oocytes regularly obtained by classical superovulation protocols. Thereafter, the authors tested the quality of these oocytes and used them for IVF, obtaining high fertilization rates (~90%), comparable to the high values regularly obtained with the new CARD methods these authors also devised recently, which have boosted the field of cryopreservation of mutant mice. Furthermore, the authors verified that the number of pups obtained after transferring all these embryos, obtained from IASe-derived oocytes and IVF into recipients, was also 2-3 times higher.

The CARD YouTube channel has also released a movie showing ampullas of oviducts from IASe-treated C57BL/6 females literally full of oocytes.

The IAS reagent used by Takeo & Nakagata is not yet commercially available. In the paper, the authors produced the IAS by themselves and titrated the product until finding the optimal dose required for maximum output. On the contrary, eCG is commercially available and is commonly used in all mouse reproductive biology and transgenic labs to promote follicle growth. Subsequent experiments will be needed to explore the validity of these results in other mouse strains and species. In addition, a commercial reliable and validated source of IAS will greatly facilitate the dissemination of this new ultra-superovulation method among the scientific community. It is also remarkable to note that the application of the IASe treatment will logically reduce the number of superovulated donor females required to obtain oocytes for cryopreservation/IVF purposes, as nicely demonstrated in this first publication, and, likely, for other aims (i.e. microinjection of DNA or RNA/genome editors to produce genetically altered mice).

Congratulations once again to Toru Takeo and Naomi Nakagata for these impressing results and for their new spectacular achievement in mouse reproductive biology!.

CARD-RPCI Cryopreservation Course Report

CARD-RPCI Cryopreservation Course Report
CARD-RPCI Cryopreservation Course Report

During the past week, 15-19 September 2014, the CARD-RPCI Mouse Sperm and Embryo Cryopreservation Course was held at Roswell Park Cancer Institute, in Buffalo, NY, USA, organized by Naomi Nakagata (CARD-Kumamoto University, Japan), Aimee Stablewski (Roswell Park Cancer Institute, Buffalo, NY, USA) and Jan Parker-Thornburg (MD Anderson Cancer Center, Houston, TX, USA). This was the second CARD course organized overseas, outside Asia, after the first course organized in Madrid in October 2013. This new practical course in North America was co-sponsored by the International Society for Transgenic Technologies (ISTT). What follows is a brief course report prepared by Aimee Stablewski and Jan Parker-Thornburg, who deserve to be praised, along with Naomi Nakagata and his CARD team, for another most successful cryopreservation course. This course report can also be downloaded from here.

CARD-­RPCI Spermand Embryo CryopreservationWorkshop
September 15-­19, 2014
Roswell Park Cancer Institute, Buffalo NY, USA

MEETING REPORT

The CARD-RPCI Sperm and Embryo Cryopreservation Workshop was recently held in Buffalo NY USA on the campus of Roswell Park Cancer Institute. Eighteen trainees, fifteen instructors and lecturers, and numerous vendors were hosted by Naomi Nakagata (Kumamoto University), Aimee Stablewski (Roswell Park Cancer Institute) and Jan Parker-Thornburg (MD Anderson Cancer Center) from September 15-19, 2014. Drs. Naomi Nakagata and Toru Takeo brought their team from CARD to assist the trainees in learning the latest techniques in embryo vitrification, sperm cryopreservation, in vitro fertilization, and the new method of vitrifying oocytes. In addition, Jorge Sztein instructed the trainees in methods of ovary freezing and subsequent implant after thaw; Lluis Montoliu provided lectures in current methods of embryo cryopreservation and CRISPR/Cas9 methods of generating genetically engineered mice, and Barbara Stone provided training in using the NSET method of embryo implant.

This was the first time that the CARD team provided training in North America, and the team (supported by Kumamoto University and Kyudo Co., LTD.) delivered a truly memorable experience for the trainees. Hands-on sessions were intense and intended to provide trainees with the knowledge and experience of performing the exacting CARD techniques. The eighteen trainees were composed of fourteen ISTT members and four non-members. Of these, three were from Europe, one from Australia, one from New Zealand, and the remainder from all parts of North America.

Hands on training included isolating unfertilized mouse oocytes, isolating and cold storage/shipping of mouse cauda epididymis, freezing/thawing mouse sperm and IVF using CARD frozen sperm and legacy sperm, fresh mouse sperm and IVF, cold stored sperm and IVF, freezing/thawing 2-cell IVF-derived mouse embryos, vitrification of mouse oocytes and embryos, IVF of vitrified mouse oocytes, ovary transplantation/ovary freezing, and embryo transfer techniques in mice (oviduct, uterus via NSET).

Didactic lectures were given on the topics of historic and scientific perspectives of embryo and sperm cryopreservation (by Jorge Sztein), comparing current embryo and sperm cryopreservation methods (by Lluis Montoliu), vitrification of oocytes and their use for IVF (by Naomi Nakagata), new US guidelines for the use of animals in research/IACUC (by Sandra Sexton), CRISPR/Cas9 and gene editing endonucleases (by Lluis Montoliu), development of database for managing mouse banking system (by Shuuji Tsuchiyama), shipping mice, frozen or refrigerated embryos/sperm across the world (by Toru Takeo) and freezing and transplantation of ovaries (by Jorge Sztein).

On Wednesday of the workshop, all of the participants were treated to a trip to Niagara Falls followed by an exceptional gala dinner (arranged by Aimee Stablewski) at a local winery. This short respite enabled the participants to replenish their energies for subsequent long days of IVF, vitrifications and surgery.

In all, it proved to be an exceptional workshop, with all participants achieving exceptional results in most, if not all, of the practicals. In fact, the depth of appreciation for being taught these methods became clear both during the closing session, where all instructors, vendors, and participants thanked Dr. Nakagata with a standing ovation, and immediately afterward, where many compliments were given. Perhaps, one of the participants said it best in an e-mail immediately following the course:

a big thank you to Prof. Nakagata, Dr. Toru Takeo and the entire CARD team. The way they have organised is unbelievable. As we all know in biological practical experiments, how much ever we take care, they never go according to plan and there is always a blooper. I must confess, I had my doubts that we would ever stick to the time table. I am glad that I was completely proven wrong. They have given attention to every bit of details and must have put lot of rehearsals behind this. They were fantastic. They answered all the questions and made sure everyone understood plus followed it up with the practicals. The Entire Card team was simply amazing and no other way to express my gratitude than simply saying “I bow to the entire CARD team”.”
Prasanna Kallingappa
University of Aukland
New Zealand

Aimee Stablewski and Jan Parker-Thornburg, as hosts, would like to acknowledge not only our lecturers, but also the CARD team who assisted the students, including Shuuji Tsuchiyama, Kiyoko Fukumoto, Yukie Haruguchi, Tomoko Kondo, Yumi Takeshita, Yuko Nakamuta, Tomoko Umeno, (all from Kumamoto University and Kyudo Co. LTD); and the CARD “adoptees”—Kristy (Kinchen) Williams (University of Florida) and Amar Dasari (Taconic). In addition, technical assistance was ably provided by Dawn Barnas (ISTT), Karstin Webber, Sandra Sexton and Leslie Curtin (all RPCI), who treated over 600 mice with hormones!

The workshop was extremely fortunate to have extremely generous vendor support, including our platinum sponsors: Charles River Laboratories (who supplied all of the mice used in the course), Leica Microsystems (who provided all of the microscopes used for the course as well as onsite support by Louise Bertrand), and Kyudo Co., LTD who provided support for the CARD instructors as well as onsite support by Nobuyuki Mikoda; our gold sponsors: the ISTT (who sponsored Lluis Montoliu’s participation and contributed to the gala dinner), Taconic (who provided instructional support from John Couse and Amar Dasari), and Regeneron (who sponsored a lecture by Lluis Montoliu); our silver sponsors: Hamilton Thorne (who provided instruction in sperm analysis and laser-assisted IVF by Nancy Mutch), IDEXX Bioresearch for course support, Paratechs for sponsoring Dr. Barbara Stone’s participation, Cell Preservation Solutions who provided course support and Lifor media for cold-storage, CosmoBio who provided course support, Transposagen who provided course support, and our bronze sponsors: Millipore EMD and mofa for equipment, Tokai-Hit for equipment, Eppendorf (Mike Bady) for course support and equipment, and VWR and Sarstedt for course support.

Finally, we thank our participants for their hard work and dedication to bring the newest transgenic technologies back to their institutions.

Aimee Stablewski, Roswell Park Cancer Institute, Buffalo, NY, USA
Jan Parker-Thornburg, MD Anderson Cancer Center, Houston, TX, USA

Transgenic Animal Technology. A Laboratory Handbook (3rd edition, 2014)

Transgenic Animal Technology. A Laboratory Handbook (3rd edition, 2014)
Transgenic Animal Technology. A Laboratory Handbook (3rd edition, 2014)

Twenty years after the publication of the first edition and twelve years after the release of the second edition of this book, Carl A. Pinkert (Auburn University, College of Veterinary Medicine, Auburn, AL, USA) in association with Elsevier, releases now the third edition of his famous transgenic manual: “Transgenic Animal Technology. A Laboratory Handbook. 3rd edition, 2014“. As it will be familiar to readers of the two previous editions of this useful and unique handbook, this is not only a manual to understand how to make a transgenic mouse. This handbook looks beyond mice and it contains protocols to prepare a wide variety of genetically-modified animals, including: rats, rabbits, poultry, fish, pigs, ruminants and non-human primates. In addition, this compilation of helpful methods includes a number of chapters devoted to understand and improve all steps of transgenesis, from vector design, analysis of transgene integration and the evaluation of transgene expression. Finally, the book also includes cryopreservation methods, an embryo culture section, a review of standard nomenclature and a selection of databases and internet resources currently available in the field.

This handbook is a worth addition to any library, laboratory or transgenic facility, complementary to other available manuals on the subject, but unique in the sense that it exquisitely illustrates current transgenic methods that can be applied to a wide variety of animal species, beyond mice.

Carl A. Pinkert has been helped in his outstanding Editorial task by a large group of co-authors, experts in their subjects, including some ISTT members: Satoshi Akagi, Anna V. Anagnostopoulos, Benjamin P. Beaton, Cory F. Brayton, Steve Brown, Anthony W.S. Chan, Tom Doetschman, Rex A. Dunham, David A. Dunn, Janan T. Eppig, Almudena Fernandez, Tatiana Flisikowska, Vasiliy Galat, Robert A. Godke, Philip Iannaccone, Michael H. Irwin, Larry W. Johnson, Yoko Kato, Teoan Kim, Alexander Kind, Bon Chul Koo, Mo Sun Kwon, Daniel J. Ledbetter, Michael J. Martin, Kazutsugu Matsukawa, Colin McKerlie, Lluis Montoliu, Paul E. Mozdziak, Akira Onishi, Paul A. Overbeek, James N. Petitte, L. Philip Sanford, Jorge A. Piedrahita, Wendy K. Pogozelski, H. Greg Polites, Edmund B. Rucker III, Marina Sansinena, Angelika Schnieke, Kumiko Takeda, James A. Thomson, Ian A. Trounce, Yukio Tsunoda, Cristina Vicente-Garcia, Kevin D. Wells, Richard N. Winn and Curtis R. Youngs.

Manipulating the Mouse Embryo. A Laboratory Manual (4th edition, 2014)

Manipulating the Mouse Embryo. A Laboratory Manual (4th edition, 2014)
Manipulating the Mouse Embryo. A Laboratory Manual (4th edition, 2014)

A bit more than 10 years later, from the last edition (3rd, 2003) published, the classical manual, the “Bible” in our field has been renewed, updated, upgraded and recently published in its fourth edition. The new “Manipulating the Mouse Embryo. A Laboratory Manual. 4th edition, 2014” has just been released, published by Cold Spring Harbor Laboratory Press and edited by Richard Behringer, Marina Gertsenstein, Kristina Vintersten Nagy and Andras Nagy. The editors, the same team that produced the 3rd edition of this useful manual, should be praised once again for a brilliant work done, accommodating the latest techniques in mouse embryo manipulation, while not forgetting the traditional procedures that must be learnt properly by anyone joining this field of animal transgenesis for the first time. The editors counted with the help of many additional collaborators, who provided materials and protocols, including many ISTT members, as Marina and Andras are, such as: Wojtek Auerbach, Ralph Brinster, Jorge Cabezas, Tracy Caroll, Lluis Montoliu, Naomi Nakagata, Jan Parker-Thornburg, Shirley Pease and Thomas Saunders, just to name a few of the many colleagues that shared their expertise and contributed to this 4th edition of the CSHLP manual.

This fourth edition is dedicated to Anne McLaren (1927-2007), one of the brave and gifted pioneer in mouse genetics and embriology, and whom do we all owe a number of standard procedures that we routinely apply in a transgenic laboratory, such as mouse embryo culturing, pseudopregnancy and embryo transfer.

The structure of this 4th edition manual follows that of the 3rd edition, in the sense that chapters and methods are grouped logically and functionally, from a very good summary on mouse genetics and mouse embryo development, to mouse colony management, and followed by a variety of main sections dealing with in vitro and in vivo work, surgical procedures, the production of transgenic mice by pronuclear microinjection, derivation of ES cell lines, using ES cells to generate germline-transmitting chimeras, genotyping protocols, parthenogenesis and nuclear transfer, assisted reproduction techniques, cryopreservation methods, techniques for visualizing gene products and finishing by a chapter devoted to setting up a micromanipulation laboratory and the usual and convenient appendix with receipts for the common buffers and solutions.

Besides this excellent updated version of the CSHLP Mouse Manual, there are several books published on this subject, including the ISTT manual (Springer, 2011) edited by Shirley Pease and Thomas Saunders, and all are worth reading and having them around for consultation, since each one brings their own contribution, their view and their solutions for the adequate learning of how to best manipulate animal embryos, and not all methods and protocols are covered in a single book, hence they are largely complementary. I will be soon reporting in this ISTT blog about additional books recently published in this field.

 

TT2014 abstracts: submission deadline is approaching (30 June)

TT2014 abstracts: submission deadline is approaching (30 June)
TT2014 abstracts: submission deadline is approaching (30 June)

From the International Society for Transgenic Technologies (ISTT) we warmly invite and encourage you all to submit your most recent and exciting results and developments in animal transgenesis to be presented at the forthcoming 12th Transgenic Technology (TT2014) meeting, which will be held in Edinburgh, Scotland, UK, on 6-8 October 2014. Deadline for submitting abstracts for the TT2014 meeting is June 30.
To submit an abstract please visit this TT2014 meeting web page.

All TT2014 participants are encouraged to submit their work as an abstract for poster presentation at the TT2014 meeting. Authors are requested to submit an abstract with the following requirements:

  • Title (max. 25 words)
  • Name authors and affiliations (first author is the presenting author).
  • Text of the communication (max. 400 words).
  • Abstracts should be submitted no later than June 30, 2014.

Accepted abstracts will be published in the scientific journal Transgenic Research (Springer), to which the ISTT is associated.

Posters
Posters will be on display in the exhibition area throughout the duration of the meeting. Poster boards are 1.00m wide x 2.00m high and we recommend posters do not exceed 1.50m in length. A supply of Velcro tabs will be available at the venue. No screws or double-sided adhesive tape will be allowed due to the damage they can cause to the boards.

Best Poster Awards
All posters presented at the TT2014 meeting will be eligible for one of the ISTT Best Poster Awards, generously sponsored by Charles River, Inc.

Oral Presentations
A limited number of abstract submissions will be selected and invited to present their findings in the form of a short oral presentation within the main meeting program. Should you be interested in being considered to speak at the meeting please select the appropriate option when submitting your abstract.

Abstracts are invited on all aspects of Transgenic Technologies, including the conference themes as listed below:

  • New technologies in animal transgenesis
  • Embryo stem cells
  • Target nucleases or Editing nucleases (ZFNs, TALENs, CRISPRs)
  • Large-scale phenotyping
  • Animal Biotechnology
  • Imaging with transgenic animals
  • Mouse models of human disease
  • Zebrafish models of human disease and transgenesis
  • Animal ethics and welfare

We are looking forward to receiving your exciting works to discuss the latest development on animal transgenesis!. See you all in Edinburgh!

Submit your abstract(s) to the TT2014 meeting in Edinburgh: 30 June

Submit your abstract(s) to the TT2014 meeting in Edinburgh: 30 June
Submit your abstract(s) to the TT2014 meeting in Edinburgh: 30 June

The submission of abstracts for the 12th Transgenic Technology (TT2014) meeting, to be held in Edinburgh (Scotland, UK), on 6-8 October, is OPEN. All TT2014 participants are encouraged to submit their work as an abstract for poster presentation at the TT2014 meeting. Authors are requested to submit an abstract with the following requirements: Title (max. 25 words), Name authors and affiliations (first author is the presenting author), and, Text of the communication (max. 400 words). Abstracts should be submitted no later than June 30, 2014. Accepted abstracts will be published in the scientific journal Transgenic Research (Springer), to which the ISTT is associated.

Posters
Posters will be on display in the exhibition area throughout the duration of the meeting. Poster boards are 1.00m wide x 2.00m high and we recommend posters do not exceed 1.50m in length. A supply of Velcro tabs will be available at the venue. No screws or double-sided adhesive tape will be allowed due to the damage they can cause to the boards. All presented Posters at the TT2014 meeting will be entitled to the Best Poster Awards, generously sponsored by Charles River.

Oral Presentations
A limited number of abstract submissions will be selected and invited to present their findings in the form of a short oral presentation within the main meeting program. Should you be interested in being considered to speak at the meeting please select the appropriate option when submitting your abstract.

Abstracts are invited on all aspects of Transgenic Technologies, including the conference themes as listed below:

  • New technologies in animal transgenesis
  • Embryo stem cells
  • Target nucleases or Editing nucleases (ZFNs, TALENs, CRISPRs)
  • Large-scale phenotyping
  • Animal Biotechnology
  • Imaging with transgenic animals
  • Mouse models of human disease
  • Zebrafish models of human disease and transgenesis
  • Animal ethics and welfare

Laboratory Course on Cryopreservation of Mouse Germplasm, Monterotondo, Italy, 20-24 October 2014

Laboratory Course on Cryopreservation of Mouse Germplasm, Monterotondo, Italy, 20-24 October 2014
Laboratory Course on Cryopreservation of Mouse Germplasm, Monterotondo, Italy, 20-24 October 2014

The Consiglio Nazionale delle Ricerche (CNR), EMMA-Monterotondo Campus International Development and The Jackson Laboratory (JAX) offer a comprehensive course on cryopreservation of mouse embryos, sperm and ovaries. This Course is financially supported by the European Union FP7 Capacities Specific Program, through the Infrafrontier-I3. The 2014 Edition of this traditional annual cryopreservation EMMA-JAX course is planned for 20-24 October 2014, organized by ISTT Member Marcello Raspa and his collaborators at CNR-EMMA Campus and with the participation of various faculty members, including Robert Taft and Jane Farley, from JAX; Ferdinando Scavizzi and Raffaele Matteoni, from CNR; Susan Marschall, from Munich; Martin Fray, from Harwell; Lluis Montoliu, from Madrid; and Kent Lloyd, from UC Davis, USA.

The 2014 edition of this course will be organized in memory of Stanley P. Leibo – Course leader 1999-2013.

The course is offered to teach methods in cryopreservation for banking of various strains of mice used in research, including inbred, transgenic and knock-out strains. Several methods of cryopreservation are now available and because no single method is adequate for all the various strains of mice being developed, a variety of methods are taught. The course is designed primarily as a “hands-on” laboratory program in which participants learn techniques for the cryopreservation of cleavage-stage embryos, spermatozoa and ovaries. Techniques include: embryo “two-step” equilibrium freezing in plastic straws, embryo non-equilibrium “ultra-rapid” cooling or “vitrification” in straws and sperm freezing and recovery of frozen sperm by in vitro fertilization. In addition, general principles of cryobiology, development of inventory databases for individual programs, and adaptation of long-term storage systems and cryogenic equipment for different situations will be presented and discussed.

Enrollment is limited to 10 participants and early application is advised. This Course is financially supported by the EU FP7 Capacities Specific Program, through the Infrafrontier-I3.

Course Fees (Fees do not include participant’s travel and lodging):

European Union’s and Associated Countries’ nationals:

  • Academic or Non-Profit Institutions: Euro 400.00
  • Other Institutions: Euro 950.00

Other Countries’ nationals:

  • Academic or Non-Profit Institutions: Euro 900.00
  • Other Institutions: Euro 1,200.00

Deadline for applications: September 10, 2014
Download here the Course application form

Course information:
Marcello Raspa, Ferdinando Scavizzi, Raffaele Matteoni
Consiglio Nazionale delle Ricerche
EMMA-Monterotondo Campus International Development
INFRAFRONTIER I3
e-mail: mraspa@emma.cnr.it

Course secretariat:
Giuliana Boera, Teresa Cuccurullo
Consiglio Nazionale delle Ricerche
EMMA-Monterotondo Campus International Development
INFRAFRONTIER I3
e-mail: cuccurullo@ibc.cnr.it