Maternal spindle transfer overcomes embryo developmental arrest caused by ooplasmic defects in mice

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Abstract

The developmental potential of early embryos is mainly dictated by the quality of the oocyte. Here, we explore the utility of the maternal spindle transfer (MST) technique as a reproductive approach to enhance oocyte developmental competence. Our proof-of-concept experiments show that replacement of the entire cytoplasm of oocytes from a sensitive mouse strain overcomes massive embryo developmental arrest characteristic of non-manipulated oocytes. Genetic analysis confirmed minimal carryover of mtDNA following MST. Resulting mice showed low heteroplasmy levels in multiple organs at adult age, normal histology and fertility. Mice were followed for five generations (F5), revealing that heteroplasmy was reduced in F2 mice and was undetectable in the subsequent generations. This pre-clinical model demonstrates the high efficiency and potential of the MST technique, not only to prevent the transmission of mtDNA mutations, but also as a new potential treatment for patients with certain forms of infertility refractory to current clinical strategies.

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First foal born in Spain using a procedure which enhances reproductive efficiency and genetic preservation in elite animals

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The first horse conceived as a result of a combination of ovum pick up (OPU) and intracytoplasmic sperm injection (ICSI), two in vitro fertilisation techniques which enhance fertility and enable the genetic preservation of elite animals, has just been born in Catalonia.
• This birth points to the future of equine assisted reproduction in Spain. It has been made possible by a project carried out entirely in the country led by the Embryotools centre based in the Barcelona Science Park in partnership with CREA Veterinaris and Embriones Equino.
• ICSI was developed in 1992 to treat men with fertility problems and is now a routine procedure in human assisted reproduction laboratories. Nonetheless, in horses there are very few laboratories (around five worldwide) that have achieved successful results.
• In 2009, the Embryotools team successfully cloned the first animals in Spain, while in 2012 the centre’s scientists were the first in Europe to achieve the birth of healthy horses after embryonic sex selection.

Barcelona, 2 June 2020. The Spanish Embryotools centre, which is based in the Barcelona Science Park (Parc Científic de Barcelona, PCB) and a pioneer in human and animal assisted reproduction, and CREA Veterinaris, which has over 25 years of experience in equine assisted reproduction, have achieved this first birth in Spain by combining ovum pick up (OPU) and intracytoplasmic sperm injection (ICSI). The foal was born on a farm in Viladrau, northern Catalonia, in the early hours of 22 May after 11 months of normal gestation.
This birth is a milestone in the future of equine assisted reproduction in Spain. It was made possible by a collaboration project between Embryotools and CREA Veterinaris in Taradell (Barcelona) along with the Seville-based firm Embriones Equinos, a leading provider of brood mares.
ICSI first emerged in 1992 for treating men with fertility problems and is now a routine procedure in human assisted reproduction laboratories. Nonetheless, in horses there are still very few laboratories (around five worldwide) that have achieved successful results. Due to the specificity of the equine oocyte, we have had to modify the procedure and use a sperm microinjection system with a Piezo system in order to achieve better results while also adapting the in vitro culture system,” says Dr Gloria Calderón, Director of Embryotools.
The process posed other problems specific to the equine species. « The first is that the mares do not respond to superovulation treatments, so we cannot get several mature ova. The second is that ultrasound-guided retrieval of ova by OPU is not easy. Although the mare copes with this procedure very well, it calls for experienced veterinarians and a good deal of training,” points out Dr Guillem Formiguera, co-director of CREA Veterinaris.
« The ova were obtained at CREA Veterinaris’ reproduction centre in Viladrau in northern Catalonia. They were then sent to our laboratory where they were matured in vitro and fertilised by means of Piezo-ICSI. The resulting embryos were cultured for eight days and sent back to the farm to be transferred to the recipient mothers, » adds Dr Nuno Costa-Borges, Scientific Director at Embryotools.

A new era of assisted reproduction in horses

One of the great advantages of the OPU-ICSI technique is that only one sperm per ovum is needed. By contrast, in conventional insemination several million spermatozoa are required for one of them to get through the barriers of the ovum, penetrate it and generate an embryo.
Most mares have their reproductive cycle during the spring and summer (the breeding season) in the months of greatest light, and few are reproductively active during the winter. So as Dr Sara Valls, co-director of CREA Veterinaris, notes: “With this technique, reproductive cycles can be extended to times when mares do not habitually cycle and vitrified embryos can be saved for transfer at a later date, as is done with embryos resulting from in vivo insemination programmes. »
This technique also allows embryos to be obtained from mares with fertility problems, and instead of one embryo per cycle, up to five can be obtained per mare every 14 days. “Although we get a high level of efficiency with the transfer programme with embryos generated by in vivo insemination, we saw that with this technique we could get embryos from stallions or mares with fertility problems which we could not do with traditional methods. So we teamed up with Embryotools to deliver this service. ICSI is so efficient at present that it can even be used to produce healthy female embryos, thus maximising the use of semen, » says Dr Formiguera.
The method enhances genetic quality in stud farms because it makes it possible to get foals from the best mares while also avoiding gestation fatigue of mares taking part in equestrian competitions. The technique can be used with any equine breed. However, it will be most sought after by breeders of high genetic value horses who will now be able to use it in Spain and will no longer have to go abroad. This IVF procedure also has great potential in terms of improvement programmes for Spanish purebred horses and elite animals. These techniques are currently in use with demand growing year after year for breeding sports horses in Europe and, albeit to a lesser extent but also rising, for other types of breeds such as the Arabian purebred and quarter horse. Scientists think that this technique also has the potential to recover other wild or endangered species.

About Embryotools
With over 30 years of experience in clinical embryology and human and animal assisted reproduction, Embryotools is a leading centre in this field in the domestic and international market. It was founded by two scientists, Gloria Calderón and Nuno Costa-Borges, both embryologists, who are driven by innovation and excellence and are passionate about the continuous improvement of assisted reproduction techniques.
In 1984, Dr Calderón was a member of the team that achieved the first pregnancy by in vitro fertilisation in Spain, and in 2009, along with Dr Costa-Borges, of the team that first successfully cloned animals in Spain. Recently, in 2019, they achieved the birth of the world’s first baby using a mitochondrial replacement technique in oocytes with poor oocyte quality.
Embryotools’ operations are divided into four major business areas: Quality Control Services to test all types of materials, culture mediums and products used in IVF laboratories; a Training Centre for sharing its experience and knowledge with professionals seeking excellence; independent Scientific and Clinical Advice and Consultancy Services, and an Animal Reproduction Centre, where the most innovative techniques are used to improve the reproductive output of elite animals. ________________________________________________________________________________________ More information: Azucena Berea • Press Officer • Barcelona Science Park • Tel. (+34) 93 403 46 62 • aberea@pcb.ub.c

Patients recruiting phase for the clinical pilot trial on maternal spindle transfer conducted by Embryotools and the Institute of Life is now concluded

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Embryotools and the Institute of Life communicate that a total of 25 patients have been successfully recruited to participate in the first registered pilot trial that is being conducted by Embryotools and the Institute of Life in Greece. The pilot trial has been approved in accordance to the terms and conditions of the Greek Law 3305/2005 and registered in a primary clinical trial registry (ISRCTN) recognized by WHO and ICMJE.

The 25 patients have been selected following the strict inclusion criteria defined in the framework of this clinical research project, which aims to evaluate the effectiveness and safety of the maternal spindle transfer technique as a means to treat infertility problems associated to cytoplasmic dysfunctions in oocytes – which manifest by a « massive » embryo development arrest in IVF cycles with conventional techniques – as well as to prevent the transmission of mitochondrial diseases.

Embryotools and the Institute of Life will like to express their gratefulness to all patients that contacted us and manifested their support and interest in participating in this project. The project is expected to be concluded within the next months and results will be made public after the acceptance of the publications in specialized peer-reviewed journals.

http://www.isrctn.com/ISRCTN11455145

 

Second baby born using maternal spindle transfer as part of the clinical research project conducted by Embryotools and the Institute of Life

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A second baby was born through the application of the maternal spindle transfer technique in the context of the clinical research project that is being carried out by Embryotools and the Institute of Life in Greece. In the framework of this clinical research pilot trial, the effectiveness and safety of the maternal spindle transfer technique is being investigated as a means to treat infertility problems associated to cytoplasmic dysfunctions in oocytes – which manifest by a “massive” embryo development arrest in IVF cycles with conventional techniques – as well as to prevent the transmission of mitochondrial diseases.

The infant was born at 07:00 am on November 11th, 2019, to a Greek women with a long history of IVF failures with conventional techniques. Both the mother and the infant are well. The birth of the first baby through maternal spindle transfer in the context of this specific clinical research study had taken place last April 2019. The infant is now seven months old, is being follow-up, and is in an excellent condition. All results of this research project that is expected to be concluded within the next months will be made public after the acceptance of the publications in specialized peer-reviewed journals.

About the maternal spindle transfer Project

The maternal spindle transfer method involves the replacement of the entire cytoplasm of oocytes, fully preserving the nuclear genome of the woman who wants to reproduce. The proof of principle of the procedure has been extensively explored in animal models and pre-clinically validated with human oocytes donated for research.  Currently, the technique is being applied in a very restrict and carefully selected cohort of patients in the context of an ongoing research protocol that was approved and is being conducted in accordance with the terms and conditions of the Greek Law 3305/2005. Based on current scientific data, the maternal spindle transfer method is still considered an experimental technique and is not an established infertility treatment nor a recognized method of medically assisted reproduction.

Embryotools achieves the world’s first pregnancy with a new nuclear transfer technique for treating infertility

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17.01.2019

Researchers from the Spanish centre Embryotools —with headquarters in the Barcelona Science Park (Parc Científic de Barcelona, PCB) — are participating in a landmark scientific event in Greece that will shape the future of assisted reproduction. In a pilot clinical trial conducted on women, which is being sponsored by the assisted reproduction centre Institute of Life in Athens, Spanish scientists have achieved the world’s first pregnancy that uses the Maternal Spindle Transfer technique to solve problems of infertility.

 

 

 

Investigadores de Barcelona y Atenas (Grecia) han logrado un embarazo en una mujer con problemas de infertilidad a la que se ha aplicado la técnica de Transferencia de Huso Materno (MST, en sus …
www.lavanguardia.com

 

Investigadores de Barcelona y Atenas. Una mujer con problemas de fertilidad logra quedarse embarazada gracias a una avanzada técnica. Este es el primer embarazo registrado en el mundo con una …
www.antena3.com

 

Collaboration EMBRYOTOOLS-MICROPTIC

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The collaboration between Embryotools and Microptic begins with the objective of performing sperm analysis in our laboratory and providing training on CASA systems (Computer Aided Semen Analysis).

We understand that andrology is a vitally important specialty in assisted reproduction units. More specifically, if we focus on the analysis of semen, applying an appropriate protocol it is possible to arrive at the diagnosis of male sterility and thus be able to offer treatments. On the other hand, sperm tests are always helpful before IVF-ICSI treatments.

 

Microptic, specialized in the analysis of semen for more than 20 years, is one of the pioneering companies in CASA systems with the SCA – Sperm Class Analyzer ®. In addition, this company already provides training and practical workshops of different levels to professionals and distributors of the sector.

Unlike manual analysis, a CASA system increases the competitiveness of laboratories thanks to the standardization of the analysis and the recording of results.

 

CASA SCA system, by Microptic

Thanks to this automatic measurement software for the analysis of sperm samples, the following parameters can be analyzed, following the WHO criteria:

Research and Development

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Our R&D centre receive support from CDTI to study new strategies to prevent prevalence of chromosomal abnormalities

Embryotools is currently involved in several research projects that aim to develop new technologies for the treatment of infertility problems.

Among them we highlight the project financed by the European Reginal Development funds (ERDF) Ref. RD 15-1-0011 that aims to develop a new technique for meiotic spindle replacement in oocytes affected by mitochondrial or other cytoplasmic problems.

Project title: Desenvolupament d’una novedosa tècnica de TRA: transferència de fus matern (MST). RD 15-1-0011.

Maternal spindle transfer overcomes embryo developmental arrest caused by ooplasmic defects in mice

Fourth Baby Born in 14 Months Using the Maternal Spindle Transfer Method as Part of Pilot Trial Conducted by the Institute of Life and Embryotools Scientific Team

 

  

Meet our team

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Founders
Nuno Costa-Borges, PhD
CEO&CSO, Co-Fondateur

Dr. Nuno Costa-Borges is an embryologist that has been involved in both research and clinical practice for over 15 years. As a PhD fellow of the Portuguese national funding agency for science, Nuno spent several years developing strategies to improve the efficiency of animal cloning, having accomplished the first animals successfully cloned in Spain in 2009. After finishing his PhD at the University Autonomous of Barcelona, he joined the embryology team of IVI Barcelona, where he has worked as a clinical embryologist. Later on, he co-founded Embryotools, where he has been devoted on offering training and consulting services to the IVF community worldwide. Additionally, as scientific director of Embryotools, Nuno has also been committed to the development of new IVF techniques, such as, the flicking method for the biopsy of blastocysts or spindle transfer for mitochondrial replacement in oocytes and embryos. Nuno’s work has led to several peer-reviewed publications and he has been invited speaker in numerous international conferences and scientific meetings, as well as awarded with prestigious prizes and grants.

Gloria Calderón, PhD
Former president

Dr. Gloria Calderón is a clinical embryologist who has been part of the team that managed to obtain the first pregnancies by IVF in Spain. With more than 30 years of experience in the field of assisted reproduction, Dr. Calderón is an international reference with extensive knowledge in clinical embryology, quality control and design of IVF laboratories. She has been the main internal scientific advisor for the IVI group between 2000 and 2013 (> 25,000 cycles / year) and is currently a guest lecturer in Master's programs at 6 Universities in Spain. Throughout her career she has provided consulting services throughout the world, and is frequently invited as speaker to the most important conferences of the sector in Europe, America, Asia, Africa and Middle East. She is the author of more than 200 publications and abstracts in international journals and conferences, including Human Reproduction, Fertility and Sterility, ESHRE, ASRM, etc. and a member of the Human Reproduction Medicine Review Committee. In addition, Dr. Calderón is part of the panel of reviewers of reputed scientific journals.

Executive board

 

Nuno Costa-Borges
CEO&CSO, Co-fondateur
Elena Chuy
Deputy CEO
David Raga
CFO

Laboratory staff

 

Enric Mestres, PhD
Directeur de laboratoire
Responsable qualité

Embryology laboratory

Mònica Acacio, MSc
Directrice du centre
vétérinaire
et de recherche

Embryology laboratory

Queralt Matia, MSc
Directrice du centre des produits

Embryology laboratory

Carolina Castelló, BSc
Directeur de la formation
Coordinatrice du centre de consultation clinique

Embryology laboratory

Andrea Villamar, Tech
Co-Directrice du centre de contrôle de la qualité

Quality control laboratory

Albert Martínez, MSc
Co-Directeur du centre de contrôle de la qualité

Quality control laboratory

Aina Franco, MSc
Coordinatrice de laboratoire

 

Quality control laboratory

Pau Soler, Tech
Technicien en embryologie

Quality control laboratory

Carlota Gutiérrez, MSc
Coordinatrice marketing

 

Embryology Laboratory

Sonia Gago, MSc
Doctorante

Embryology laboratory

Aran Santamaría, MSc
Technicienne en embryologie

Quality control laboratory

Alba González, MSc
Technicienne en embryologie

Quality control laboratory

 

Administration

 

Lara Pérez
Finance manager & Quality manager
Maria Campos
Operations manager & Quality manager
Karen Mitma
Accountig assistant
Stephanie Melega
Office manager
Carles Ortega Ragull
Software developer manager

External advisors

 

Dagan Wells, PhD
Conseiller scientifique

Dagan Wells has been actively involved in preimplantation genetic diagnosis (PGD) and the study of human gametes and embryos for over two decades, conducting his first PGD cases in 1992. He spent several years developing novel PGD tests at University College London, accomplishing the first comprehensive chromosome analysis of cells from human embryos in 1998, using a combination of whole genome amplification and comparative genomic hybridisation (CGH). In 1999 Dagan moved to the United States and joined Reprogenetics, one of the world’s largest providers of PGD services. In 2003 he initiated Reprogenetics’ highly successful single gene PGD program, testing embryos for numerous serious inherited conditions. Dagan later joined the faculty of Yale University Medical School, where he spent four years running a research laboratory, before returning to the UK in late 2007. His research group is now located in the Nuffield Department of Obstetrics and Gynaecology at the University of Oxford. Dagan’s work has led to the publication of more than 120 peer-review publications and book chapters and has been recognized with the award of numerous prizes and grants. He is a Fellow of the Royal College of Pathologists and currently serves on the Editorial Boards of several international journals. Dagan also directs Reprogenetics-UK, an independent laboratory offering state-of-the-art PGD services to IVF clinics throughout the UK and internationally.

Klaus Rink, PhD
Conseiller scientifique

German physicist, specialized in experimental physics, fluent in English and French. He is renowned for research and development of laser products for medical applications in the fields of in vitro fertilization, tissue welding and the treatment of kidney stones. In 2000 he co-founded OCTAX Microscience GmbH in Germany. OCTAX develops and manufactures IVF medical equipment, ranging from therapeutic to diagnostics appliances. OCTAX is reputed worldwide for producing the most reliable and accurate laser equipment, researched and developed by Dr. Rink since 1992. Scientific achievements: /1st birth after laser assisted hatching (1.48 μm) /1st birth after laser assisted blastomere biopsy and PGD /1st birth after laser assisted polar body biopsy /1st birth after laser assisted immobilization of spermatozoa /1st pregnancy after laser assisted zona drilling for ICSI /1st pregnancy after laser assisted removal of necrotic blastomeres from cryopreserved embryos

Gabriel Clérico
Vétérinaire

Gabriel José Clérico is a distinguished Argentinian biotechnologist specializing in equine reproductive technologies. He has played a pivotal role in advancing assisted reproduction in horses, particularly in the areas of oocyte vitrification and intracytoplasmic sperm injection (ICSI). In a landmark study published in Theriogenology in 2021, Clérico and his team reported the successful birth of live foals from vitrified/warmed immature equine oocytes matured with or without melatonin supplementation. This achievement marked a significant milestone in equine reproductive science, demonstrating the viability of vitrified oocytes in producing healthy offspring. Beyond his groundbreaking work in oocyte vitrification, Clérico has contributed to the broader field of equine assisted reproduction through studies on oocyte maturation dynamics and the timing of polar body extrusion, further informing best practices in ICSI procedures. Gabriel Clérico's contributions have not only advanced scientific understanding but have also had practical implications for equine breeding programs, offering new avenues for preserving and propagating valuable genetic lines.  

Xavier Santamaria, M.D., PhD
Conseiller médico-scientifique

Board Ob/Gyn specialist in Reproductive Medicine. He obtained a post-graduate fellowship in Reproductive Endocrinology and Infertility at Yale University (USA) and completed his Ph.D. at the Universitat Autònoma Barcelona (Spain). Dr. Santamaria war the Director of the International Department at the IVI between 2012 and 2016. In 2016, he joined Dr. Simon's Research group as a Senior Principal Investigator, where he has developed his research related to Asherman's Syndrome. His mail clinical and scientific interest is in understanding the regenerative capacity of the human endometrium and developing strategies to regenerate the endometrium to improve embryo implantation. His group was the first to complete a trial using human bone marrow stem cells to treat Asherman's syndrome and endometrial atrophy (PMDI:27005892) demonstrating this approach's feasibility in treating endometrial pathologies. As a result, CD133+ cells have been designed as the first Orphan Drug (ODD) by the European Medicines Agency (EMA) and Food and Drug Administration (FDA) in the field of gynecological research. Dr. Santamaria has also participated as an inventor in 7 different patent applications, and is a co-founder and scientific director of 4 different start-up companies. He has published 27 papers in peer-reviewed journals (H-index of 15) and 12 book chapters. He has been an invited speaker at more than 70 international conferences and was awarded the prize for Best Oral Presentation for Young Investigators at the Society of Ginecological Investigation Meeting in 2009. He has directed one doctoral thesis.

Olmo Rigoni
Conseiller commercial

Legal Advisors

Guillermo Gilabert Marqués
Associé codirecteur du cabinet d’avocats Manubens

Practice Areas: Corporate Law./ Sectors: Biotechnology/Family Businesses/Foundations and Non-Profit Organisations/New Technologies/Intellectual Property/ Experience: Design of legal and business structures; formalisation of corporate agreements and conflict management. Commercial and competition law. Foreign investment. Public procurement. Legal advice for foundations and non-profit organisations. Corporate and financial law. Law applied to new technologies, biotechnology and renewable energies. Legal services for several companies operating in different sectors, both at a national and international level./ Qualifications: Law degree, Universitat de Valencia. Lecturer on the Master's Degrees: Programme of Owner Managers and Programme for Management Developement, ESADE. Lecturer in Corporate Law ESCODI and Universitat de Barcelona.

Antonio Jesús Jimena Hidalgo
Avocat du cabinet d’avocats Manubens

Practice Areas: Corporate Law./ Sectors: Biotechnology/Family Businesses/Foundations and Non-Profit Organisations/New Technologies/Intellectual Property/ Experience: Design of legal and business structures; formalisation of corporate agreements and conflict management. Commercial and competition law. Foreign investment. Public procurement. Legal advice for foundations and non-profit organisations. Corporate and financial law. Law applied to new technologies, biotechnology and renewable energies. Legal services for several companies operating in different sectors, both at a national and international level./ Qualifications: Law degree, Universitat de Málaga. xxxxxxxx xx xxxt xxxxxxxx xxxxxxxx xxxxxxxxx xx xxxxx xxxxxxxx xxx xxxxxxxxx xxx xxxxxxxxxx xxxxxxxxxxxx, XXXXX. Xxxxxxxx xx Xxxxxxxxx Xxx XXXXXX xxx Xxxxxxxxxxx xx Xxxxxxxxx