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. 

 

Unlike conventional cryobanks, which primarily store genetic information in the form of non-viable biological samples, we extend this concept through advanced cryopreservation techniques, cellular methodologies, and state-of-the-art stem cell technologies to include living biological material. This effort is supported by continuous access to wildlife samples through the zoo community, the Leibniz-IZW Pathology Unit (Department of Wildlife Diseases), and several major research initiatives, including Frozen Dumbo, EUROVA, and naked mole-rat research projects.

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

The cryobank contains tissue samples from more than 500 species (approximately 600 taxa), which can be used for the establishment of cell cultures as well as for genetic, physiological, and evolutionary research. In addition, the collection includes approximately 10,000 tissue samples from naked mole-rats originating from both wild populations and the Leibniz-IZW breeding colony. Due to their exceptional longevity, cancer resistance, and tolerance to hypoxia, naked mole-rats represent a valuable model organism for ageing research.

2) Fibroblast culture – for generating iPSC

The department establishes and maintains fibroblast cell lines from approximately 350 taxa representing all major vertebrate groups. These include numerous threatened species, as well as species that are extinct in the wild, such as the northern white rhinoceros. The collection provides the foundation for generating induced pluripotent stem cells (iPSCs) and will be strategically expanded to include key conservation species such as rhinoceroses, elephants, and wolves.

A major component of this effort is the Israel-German Ark of Life (IGAL), established in 2017 in collaboration with the Ramat Gan Safari Park, Israel. Through the zoo’s wildlife hospital, which treats approximately 4,000 animals annually, tissues and cell cultures from an exceptionally diverse range of species—including many rare migratory birds—are secured and preserved within the cryobank.

3) Gametes, Embryos, ESC

Since the 1990s, the department has collected and maintained cryopreserved sperm samples from approximately 100 species. The value of such genetic resource banks has been demonstrated through their successful application in artificial insemination and in vitro fertilization programs for elephants and rhinoceroses.

In parallel, the department develops novel assisted reproductive technologies (ART) to improve the cryopreservation of gametes, reproductive tissues, and embryos, as well as in vitro and in vivo fertilization and embryo transfer procedures. Leibniz-IZW is internationally recognized as a leader in the development of species-specific reproductive technologies. This expertise is reflected in patents related to elephant artificial insemination and rhinoceros oocyte retrieval, as well as in successful giant panda breeding programs in Berlin and Chengdu.

The collection is integrated into national and international biobanking networks, including the Frozen Zoo (San Diego), the Central Biobank of Charité, the German Biobank Node (GBN), the Cryopreservation Unit W430 of the German Cancer Research Center (DKFZ), the Interdisciplinary Center for Biobanking Lübeck (ICB-L), and the European, Middle Eastern and African Society for Biopreservation and Biobanking (ESBB). Additional support is provided through the Leibniz Association (SAW programme) for naked mole-rat research, the German Community of Cryobanks (GDK), and the BMFTR-funded BioRescue project. A unique feature of the collection is its extensive archive of fibroblast cultures derived from mammals, birds, reptiles, and amphibians, representing one of the most comprehensive wildlife cell repositories in Europe.

Looking ahead, the Bio-Cryobank will be transformed into a fully digitalized and largely automated infrastructure. Since 2021, an additional cell culture laboratory has been available for specialized applications, including the cultivation of amphibian cell lines. Furthermore, the construction of a new Cell Biology Building is currently being planned.

Hildebrandt TB, Holtze S, Biasetti P, Bohner J, Cizmar D, Colleoni S, de Mori B, Diecke S, Galli C, Hayashi K, Kangethe F, Korody ML, Lekolool I, Ndereeh D, Kariuki L, Lazzari G, Mijele D, Mutisya S, Ngulu S, Omondi P, Seet S, Zywitza V, Stejskal J, Goeritz F (2026): De-extinction of the Northern White Rhinoceros (Ceratotherium simum cottoni). J Reprod Dev 72: 87–94, 2026. https://doi.org/10.1262/jrd.2025-122

Phan NT, Uldry J, Matera-Witkiewicz A, De Wilde A (authors), Schrade L, Stumptner C, Escámez T, Gleńska-Olender J, Foesel B (key contributors)(2026). Quality Handbook for Biobanks (QHB). Zenodo. https://doi.org/10.5281/ZENODO.18197174

Geltinger F, Holtze S, Hildebrandt TB, Ochs M, Brandenberger C (2025): Stereological analysis of the naked mole-rat Heterocephalus glaber provides evidence of altered surfactant function and late alveolarization. Am J Physiol Lung Cell Mol Physiol. 2025 Jul 1;329(1):L84-L96, https://doi:10.1152/ajplung.00246.2024

Holtze S, Demirtürk D, Ohlenschläger O, Hegele S, Siniuk K, Förste S, Raghavan V, Groth M, Bens M, Hasse N, Martel A, Lukač M, Cizelj I, Fischer M, Liu H, Hildebrandt TB, Hoffmann S, Sahm A (2025): The transcriptome of the olm provides insights into its evolution and gene expression. Scientific Reports 15:28324. https://doi.org/10.1038/s41598-025-10073-3

Biasetti P, Mercugliano E, Schrade L, Spiriti MM, Goeritz F, Holtze S, Seet S, Galli C, Stejskal J, Colleoni S, Cizmar D, Simone R, Hildebrandt TB, de Mori B (2024): Ethical assessment of genome resource banking (GRB) in wildlife conservation. CRYOBIOLOGY 117, https://doi.org/10.1016/j.cryobiol.2024.104956

Subiran Adrados C, Ørnes Olesen H, Vernimmen Olesen S, Pors SE, Holtze S, Hildebrandt TB, Andersen CY, Kristensen SG (2024): Exploring the effect of PRP on vascularization and survival of follicles in human ovarian tissue transplanted to immunodeficient mice. Reproductive BioMedicine Online, S1472-6483(24)00463-2. https://doi.org/10.1016/j.rbmo.2024.104274

Simone R, Cizmar D, Holtze S, Mulot B, Lamglait B, Knauf-Witzens T, Weigold A, Hermes R, Hildebrandt TB (2024): Cryopreservation of okapi (Okapia johnstoni) oocytes following in vitro maturation. Theriogenology Wild 4 (2024) 100088. https://doi.org/10.1016/j.therwi.2024.100088

Schrade L, Mah N, Bandrowski A, Chen Y, Dewender J, Diecke S, Hiepen C, Lancaster MA, Marques-Bonet T, Martinez S, Mueller SC, Navara C, Prigione A, Seltmann S, Sochacki J, Sutcliffe MA, Zywitza V, Hildebrandt TB, Kurtz A (2024): A Standardized Nomenclature Design for Systematic Referencing and Identification of Animal Cellular Material. Animals 2024, 14, 1541. https://doi.org/10.3390/ani14111541

Ruiz-Orera J, Miller DC, Greiner J, Genehr C, Grammatikaki A, Blachut S, Mbebi J, Patone G, Myronova A, Adami E, Dewani N, Liang N, Hummel O, Muecke MB, Hildebrandt TB, Fritsch G, Schrade L, Zimmermann WH, Kondova I, Diecke S, van Heesch S, Hübner N (2024): Evolution of translational control and the emergence of genes and open reading frames in human and non-human primate hearts. Nat Cardiovasc Res 3, 1217–1235. https://doi.org/10.1038/s44161-024-00544-7

Hutchinson A, Appeltant R, Burdon T, Bao Q, Bargaje R, Bodnar A, Chambers S, Comizzoli P, Cook L, Endo Y, Harman B, Hayashi K, Hildebrandt TB, Korody ML, Lakshmipathy U, Loring JF, Munger C, Ng A, Novak B, Onuma M, Ord S, Paris M, Pask A, Pelegri F, Pera M, Phelan R, Rosental B, Ryder OA, Sukparangsi W, Sullivan G, Tay NL, Traylor-Knowles N, Walker S, Weberling A, Whitworth DJ, Williams SA, Wojtusik J, Wu J, Ying Q-L, Zwaka TP, Kohler TN (2024): Advancing stem cell technologies for conservation of wildlife biodiversity. Development 151, dev203116. https://doi.org/10.1242/dev.203116

Hildebrandt TB and Holtze S (2024): Advanced assisted reproduction technologies in endangered mammalian species. Reprod Dom Anim. 59(Suppl. 3):e14700. https://doi.org/10.1111/rda.14700

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.

Hildebrandt TB, Holtze S, Colleoni S, Hermes R, Stejskal J, Lekolool I, Ndeereh D, Omondi P, Kariuki L, Mijele D, Mutisya S, Ngulu S, Diecke S, Hayashi K, Lazzari G, de Mori B, Biasetti P, Quaggio A, Galli C, Goeritz F (2023): In vitro fertilization program in white rhinoceros. Reproduction 166(6): 383–399. https://doi.org/10.1530/REP-23-0087.

Gerhardt P, Begall S, Frädrich C, Renko K, Hildebrandt TB, Holtze S, Heinrich A, Sahm A, Meci X, Köhrle J, Rijntjes E, Henning Y (2023): Comparative analysis of thyroid hormone systems in rodents with subterranean lifestyle. Scientific Reports 13:3122 (2023). https://doi.org/10.1038/s41598-023-30179-w

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. https://doi.org/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. https://doi.org/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. https://doi.org/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. https://doi.org/10.1016/j.theriogenology.2021.04.006

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. https://doi.org/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. https://doi.org/10.1071/RD20265

Hermes R, Hildebrandt TB, Göritz F, Fasel NJ, Holtze S (2019). First cryopreservation of phyllostomid bat sperm. Theriogenology. 131:28-31. https://doi.org/10.1016/j.theriogenology.2019.03.014

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. https://doi.org/10.1016/j.bone.2019.115035

Shekhidem AH, 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. https://doi.org/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. https://doi.org/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. https://doi.org/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. https://doi.org/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. https://doi.org/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. https://doi.org/10.1371/journal.pgen.1007272

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. https://doi.org/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. https://doi.org/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. https://doi.org/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. https://doi.org/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. https://doi.org/10.1007/s10344-014-0858-4

Arav A, Saragusty J (2014): Directional freezing of spermatozoa and embryos. Reprod Fertil Dev 26, 83-90. https://doi.org/10.1071/Rd13295