Towards the next level of biobanking

While biodiversity is decreasing at an alarming rate, cryobanks contribute to preserving the ‘library of life‘. Our large-scale bio-cryobank, including not only tissues, but also gametes, fibroblasts, induced pluripotent stem cells, embryonal stem cells and embryos, is planned to be upgraded to an automated, fully digitized system. 

Project details
Duration: 01/2013 – 12/2025
Third-party funded: yes
Involved Department(s): Dept Reproduction Management
Leibniz-IZW Project Leader(s): Thomas Hildebrandt (Dept Reproduction Management)
Leibniz-IZW Project Team:

Frank Göritz, Robert Hermes, Susanne Holtze, Daniel Čižmár, Charlotte Okolo, Nadia Kirchler, Jette Ziep, Friederike Rauch (all: Dept Reproduction Management)
Consortium Partner(s):

Dr. Sebastian Diecke, Core Facility Stammzellen, Max Delbrück Center for Molecular Medicine (MDC), Berlin, Germany

Prof. Cesare Galli, AVANTEA Laboratory of Reproductive Technologies, Cremona, Italy

Prof. Katsuhiko Hayashi, Department of Stem Cell Biology and Medicine, Kyushu University, Japan

Prof. Dr. Johannes Schenkel, Chairman of Gemeinschaft Deutscher Kryobanken e.V. (GDK), Cryopreservation Unit W430, German Cancer Research Centre (DKFZ) / Institute of Physiology and Pathophysiology, Heidelberg University

Prof. Dr. Michael Hummel, Central Biobank Charité, coordinator of the German Biobank Node (GBN), Charité Universitätsmedizin Berlin, Germany

Professor Dr. Jens K. Habermann, Director of the Interdisciplinary Center for Biobanking-Lübeck (ICB-L), President of ESBB (European, Middle Eastern and African Society for Biopreservation and Biobanking), University of Lübeck, Germany

Dr. Oliver Ryder, Frozen Zoo, San Diego Global, USA
Current Funding Organisation: German Federal Ministry of Education and Research (BMBF), Leibniz Association (SAS-2016-2020-IZW-LFV; SAW-2018-DRFZ)
Research Foci:
Understanding traits and evolutionary adaptations
Improving population viability
Developing theories, methods, and tools
Photo gallery

The current extinction rate during the ongoing earth’s sixth great extinction event urgently requires new conservation strategies. Besides protection of habitat and species in and ex situ, one priority is the preservation of the current ‘library of life’. Cryopreservation of gametes, embryos, tissues and specific cell lines is important to preserve and maintain biodiversity of endangered species.

Whereas the majority of existent cryobanks aim at preserving mainly genetic material or other non-alive items, we extend this classical concept by including also living materials, involving the use of improved cryopreservation techniques, cellular methods and state-of-the-art stem cell technologies. Based on the long-term access to wildlife samples provided by stakeholders from the zoo community, the IZW pathology unit (Department Wildlife Diseases) as well as several projects involving free-ranging wildlife (e.g. Frozen Dumbo), we continually expand this bio-insurance – ‘bio-cryobank’.

Comprising tissues, fibroblast cultures, gametes, embryonal stem cells (ESC), induced embryonal stem cells (iESC), and embryos, it is embedded in the efforts of several further large collections, such as the Frozen Zoo in San Diego, Central Biobank Charité, German Biobank Node (GBN), Cryopreservation Unit W430 of the German Cancer Research Center, Interdisciplinary Center for Biobanking-Lübeck (ICB-L), and the European, Middle Eastern and African Society for Biopreservation and Biobanking (ESBB). Among funding from the Leibniz Association (SAW) for naked mole rat research, the Community of German Cryobanks (GDK) for the project “White Rhinoceros”, we receive financial support from the BMBF-funded project BioRescue. A European unique selling point of our biobank is the extensive archive of fibroblast cultures of bird, reptile and amphibian species in addition to mammalian cells.

This valuable, living sample material could be used also for example to study receptors in regard of their Covid-19 susceptibility across the different mammal species.

1) Tissue – cell culture, genetic studies and basic research

Besides tissues stored for the future initiation of cell culture and for genetic and physiological research, we also maintain an extensive library of ~1000 samples of various naked mole rats tissues both from captivity and the wild for basic research; naked mole rats display a unique reproductive system, extraordinary cancer and hypoxia resistance, and exceptional longevity, and therefore serve as a highly interesting animal model to investigate evolutionary adaptations of healthy aging.

2) Fibroblast culture – for generating iPSC

Our department collects and manages fibroblast cell lines cultured from 118 species. Of these, according to the IUCN red list (http://www.iucnredlist.org) 26 are classified as vulnerable, 11 as endangered and 14 as critically endangered (e.g. the Spix’ Macaw); 3 are even extinct in the wild (e.g. the northern white rhinoceros). With the help of third party and future funding opportunities this existing bio-cryobank will be expanded and complemented with in vitro derived embryonic stem cells and induced pluripotent stem cells from keystone species such as rhinoceroses, elephants, and grey wolves. In 2017, the Israel-German Ark of Life (IGAL) was initiated, a project of the Ramat Gan Safari (Israel) together with the Department Reproduction Management, aiming at conserving tissues and cell cultures collected at the wildlife clinic of the zoo. With approximately 4000 annual patients, it offers access to a high diversity of species, including various rare migratory birds.

3) Gametes, Embryos, ESC

In addition, our department has been collecting and managing cryopreserved sperm from a total of 45 species since the 1990s. The value of such sperm banks was demonstrated by the use of elephant and rhinoceros sperm in artificial insemination (AI) and in vitro fertilization (IVF) programmes. Scientists of the Department of Reproduction Management develop new methods of assisted reproduction technologies (ART) to improve cryopreservation of gametes, reproductive tissues and embryos, and to refine in vitro and in vivo fertilization and embryo transfer techniques for a variety of species. The Leibniz-IZW is leading in developing assisted reproduction techniques tailored to specific species, as documented by several patents (elephant artificial insemination, rhinoceros ovum pickup) and by the success in breeding giant pandas at Berlin Zoo and at the Chengdu Research Base of Giant Panda Breeding in China.

The next step – with the help of future funding opportunities which will allow for the necessary investment – will be to upgrade this large-scale comprehensive wildlife bio- cryobank to an automated, fully digitised system. A second cell culture laboratory is available since 2021 for special tasks such as e.g. the culture of amphibian cell lines. Building and establishing a new "cell biology" complex at the IZW starting is in planning.

Selected Publications

  • Lázaro J, Costanzo M , Sanaki-Matsumiya M, Girardot C, Hayashi M, Hayashi K, Diecke S, Hildebrandt TB, Lazzari G, Wu J, Petkov S, Behr R, Trivedi V, Matsuda M, Ebisuya M (2023): A stem cell zoo uncovers intracellular scaling of developmental tempo across mammals. Cell Stem Cell 30: 1-12, https://doi.org/10.1016/j.stem.2023.05.014.
  • Zywitza V, Frahm S, Krüger N, Weise A, Göritz F, Hermes R, Holtze S, Colleoni S, Galli C, Drukker M, Hildebrandt TB, Diecke S. (2022). Induced pluripotent stem cells and cerebral organoids from the critically endangered Sumatran rhinoceros. iScience. 25:105414. doi: 10.1016/j.isci.2022.105414.
  • Hayashi M, Zywitza V, Naitou Y, Hamazaki N, Goeritz F, Hermes R, Holtze S, Lazzari G, Galli C, Stejskal J, Diecke S, Hildebrandt TB, Hayashi K. (2022). Robust induction of primordial germ cells of white rhinoceros on the brink of extinction. Science Advances 8:eabp9683. doi: 10.1126/sciadv.abp9683.
  • Zywitza V, Rusha E, Shaposhnikov D, Ruiz-Orera J, Telugu N, Rishko V, Hayashi M, Michel G, Wittler L, Stejskal J, Holtze S, Göritz F, Hermes R, Wang J, Izsvák Z, Colleoni S, Lazzari G, Galli C, Hildebrandt TB, Hayashi K, Diecke S, Drukker M. (2022). Naïve-like pluripotency to pave the way for saving the northern white rhinoceros from extinction. Sci Rep. 12:3100. doi: 10.1038/s41598-022-07059-w.
  • 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.
  • Del Marmol D, Holtze S, Kichler N, Sahm A, Bihin B, Bourguignon V, Dogné S, Szafranski K, Hildebrandt TB, Flamion B. (2021). Abundance and size of hyaluronan in naked mole-rat tissues and plasma. Sci Rep. 11:7951. doi: 10.1038/s41598-021-86967-9.
  • 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
  • Hermes R, Hildebrandt TB, Göritz F, Fasel NJ, Holtze S (2019). First cryopreservation of phyllostomid bat sperm. Theriogenology. 131:28-31. doi: 10.1016/j.theriogenology.2019.03.014. Epub 2019 Mar 19. PMID: 30933687
  • Carmeli-Ligati S, Shipov A, Dumont M, Holtze S, Hildebrandt TB, Shahar R (2019). The structure, composition and mechanical properties of the skeleton of the naked mole-rat (Heterocephalus glaber). Bone. 128:115035. doi: 10.1016/j.bone.2019.115035.
  • Adwan Shekhidem H, Sharvit L, Leman E, Manov I, Roichman A, Holtze S, M Huffman D, Y Cohen H, Hildebrandt TB, Shams I, Atzmon G (2019). Telomeres and Longevity: A Cause or an Effect? International journal of molecular sciences. 20(13):3233. doi: 10.3390/ijms20133233.
  • Heinze I, Bens M, Calzia E, Holtze S, Dakhovnik O, Vyssokikh M, Sahm A, Kirkpatrick JM, Szafranski K, Romanov N, Singer S, Ermolaeva M, Platzer M, Hildebrandt TB, Ori A (2018). Species comparison of liver proteomes reveals links to naked mole-rat longevity and human aging. BMC Biol. 16:82. doi: 10.1186/s12915-018-0547-y.
  • Bens M, Szafranski K, Holtze S, Sahm A, Groth M, Kestler HA, Hildebrandt TB, Platzer M (2018). Naked mole-rat transcriptome signatures of socially-suppressed sexual maturation and links of reproduction to aging. BMC Biol. 16:77. doi: 10.1186/s12915-018-0546-z.
  • 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.
  • Hermes R, Hildebrandt TB, Göritz F (2018): Cryopreservation in rhinoceros - setting a new benchmark for sperm cryosurvival. PLOS ONE 13, e0200154. doi:10.1371/journal.pone.0200154.
  • Sahm A, Bens M, Szafranski K, Holtze S, Groth M, Görlach M, Calkhoven C, Müller C, Schwab M, Kestler HA, Cellerino A, Burda H, Hildebrandt TB, Dammann P, Platzer M (2018). Long-lived rodents reveal signatures of positive selection in genes associated with lifespan. PLoS genetics 14.3: e1007272.
  • Debebe T, Biagi E, Soverini M, Holtze S, Hildebrandt TB, Birkemeyer C, Wyohannis D, Lemma A, Brigidi P, Savkovic V, König B, Candela M, Birkenmeier G (2017). Unraveling the gut microbiome of the long-lived naked mole-rat. Sci Rep. 7:9590. doi: 10.1038/s41598-017-10287-0.
  • 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.
  • Saragusty J, Osmers J-H, Hildebrandt TB (2016): Controlled ice nucleation - is it really needed for large-volume sperm cryopreservation? THERIOGENOLOGY 85, 1328-1333. doi:10.1016/j.theriogenology.2015.12.019.
  • Arav A, Saragusty J (2016): Directional freezing of sperm and associated derived technologies. ANIM REPROD SCI 169, 6-13. doi:10.1016/j.anireprosci.2016.02.007.
  • Prieto MT, Sanchez-Calabuig MJ, Hildebrandt TB, Santiago-Moreno J, Saragusty J (2014): Sperm cryopreservation in wild animals. EUR J WILDL RES 60, 851-864. doi:10.1007/s10344-014-0858-4.
  • Arav A, Saragusty J (2014): Directional freezing of spermatozoa and embryos. REPROD FERTIL DEV 26, 83-90. doi:10.1071/Rd13295.