Translational research for developing assisted reproduction technologies for endangered mammals (EUROVA – European Oocyte Biology Research Innovation Training Net)

The EUROVA consortium aims at developing new techniques and methods for in vitro maturation (IVM) of oocytes, in vitro fertilization (IVF) and in vitro culture (IVC) of embryos. These may be used to advance new conservation tools for highly endangered mammals, such as the endangered rhinoceros family.

Project details
Duration: 06/2020 – 06/2022
Third-party funded: yes
Involved Department(s): Dept Reproduction Management, Dept Evolutionary Genetics
Leibniz-IZW Project Leader(s): Thomas Hildebrandt (Dept Reproduction Management), member of the ethics sub-committee of EUROVA
Leibniz-IZW Project Team:

Raffaella Simone

Consortium Partner(s):

University College Dublin (UCD), National University of Ireland (Ireland), Universita Degli Studi di Milano (Italy), Vrije Universiteit Brussel (Belgium), The Babraham Institute (UK), Euvitro SLU (Spain), Universiteit Gent (Belgium), Consejo Superior de Investigaciones Cientificas, M.P. (Spain), Cherry Biotech (France), Region Hovedstaden (Denmark)

Current Funding Organisation:

EU: Marie Skłodowska-Curie Action ETN EUROVA

PI: University College Dublin; PhD-program for altogether 15 students, total budget 4,5 Mio €; Leibniz-IZW: 1 PhD, funded with 158 k€

Research Foci:
Understanding traits and evolutionary adaptations
Understanding the environmental context
Improving population viability
Developing theories, methods, and tools
Videos about the project


In light of the Earth’s sixth great extinction event driven by man, traditional conservation strategies such as habitat protection and ex situ breeding combined with reintroduction programmes will be not sufficient to stop or even to slow down this process. Currently, 22% of the mammals are at risk of extinction ( The family Rhinocerotidae is particularly affected, with three of the five extant species listed as critically endangered (Sumatran, Javan, and black rhinoceros), one listed as vulnerable (greater one-horned rhinoceros), and one, the southern white rhinoceros listed as near threatened. The complementary cutting-edge expertise of the EUROVA consortium provides a unique opportunity for the development of new techniques for in vitro maturation (IVM) of oocytes, in vitro fertilisation (IVF) and in vitro culture (IVC) of embryos. The strategic translation of the interdisciplinary and intersectoral research into practical conservation tools would offer a blue print for saving critically endangered mammalian species from extinction. 

In our research approach the basis for the development of new techniques and methods for the conservation of species is the regular collection of rhinoceros egg cells in European zoos. The collection is performed with our recently patented collection device (DE 102017002614, US2018/0250032). We aim to carry out comparative studies on genetic, structural and functional parameters of these oocytes. Due to the value of each oocyte, non-destructive, high quality time lapse technologies (Geri+, Merck KGaA) equipped with software for computer-based event detection will provide the majority of the missing information on non-invasive biomarkers of oocyte fitness and adequate embryo development. Additionally, selective invasive investigations (e.g. transcriptome analysis) will be performed. The resulting physiological landmarks will be used to translate optimized in vitro production protocols in murine, cattle and human models into adopted protocols for endangered mammalian species.

Expected results

This project will help elucidating species-specific and general requirements for in vitro Duplication Of Maternal Environment (ivDOME) for an optimal gamete maturation and pre-implantation embryo development with the final goal for achieving reproductively healthy adult offspring.

Innovation, impact, applicative outcome(s)

The scientific results will provide a future role model for global conservation strategies rescuing critically endangered mammal species from extinction. Due to national and international conservation laws innovations regarding endangered species cannot be commercialised. However, the innovative character of the project and the substantial funding used offer high potential that equipment and/or techniques could be patented and commercially applied in areas outside conservation. The consortium scientists will secure innovations in form of patents.

Selected Publications

  • Simone R, Čižmár D, Holtze S, Michel G, Sporbert A, Okolo C, Hildebrandt TB. (2023). In vitro production of naked mole-rats’ blastocysts from non-breeding females using in vitro maturation and intracytoplasmic sperm injection. Sci Rep 13, 22355 (2023).  
  • Hildebrandt TB, Hermes R, Goeritz F, Appeltant R, Colleoni S, de Mori B, Diecke S, Drukker M, Galli C, Hayashi K, Lazzari G, Loi P, Payne J, Renfree M, Seet S, Stejskal J, Swegen A, Williams SA, Zainuddin ZZ, Holtze S. (2021). The ART of bringing extinction to a freeze - History and future of species conservation, exemplified by rhinos. Theriogenology. 169:76-88. doi: 10.1016/j.theriogenology.2021.04.006. Epub 2021 Apr 18. PMID: 33940218.
  • Hayashi K, Cesare G, Diecke S, Hildebrandt TB (2021): Artificially produced gametes in mice, humans and other species. REPROD FERTIL DEV 33, 91–101. doi:10.1071/RD20265

  • Hildebrandt TB*, Holtze S*, Biasetti P, Colleoni S, de Mori B, Diecke S, Göritz F, Hayashi K, Hayashi M, Hermes R, Kariuki L, Lazzari G, Mijele D, Mutisya S, Ndeereh D, Ngulu S, Seet S, Zwilling J, Zywitza V, Stejskal J, Galli C (2021): Conservation Research in Times of Covid-19 – the Rescue of the Northern White Rhino. J APPL ANIM ETHICS RES 1, 1-22. doi:10.1163/25889567-BJA10009

  • Hildebrandt TB, Hermes R, Colleoni S, Diecke S, Holtze S, Renfree MB, Stejskal J, Hayashi K, Drukker M, Loi P, Göritz F, Lazzari G, Galli C (2018): Embryos and embryonic stem cells from the white rhinoceros. NAT COMMUN 9, 2589. doi:10.1038/s41467-018-04959-2.

  • Saragusty J, Diecke S, Drukker M, Durrant B, Ben-Nun I, Galli C, Göritz F, Hayashi K, Hermes R, Holtze S, Johnson S, Lazzari G, Loi P, Loring JF, Okita K, Renfree MB, Seet S, Voracek T, Stejskal J, Ryder OA, Hildebrandt TB (2016): Rewinding the process of mammalian extinction. ZOO BIOL 35, 280-292. doi:10.1002/zoo.21284.