The northern white rhinoceros (NWR) is probably the earth's most endangered mammal. To rescue the functionally extinct species, we aim to employ induced pluripotent stem cells (iPSCs) to generate gametes and subsequently embryos in vitro. To elucidate the regulation of pluripotency and differentiation of NWR PSCs, we generated iPSCs from a deceased NWR female using episomal reprogramming, and observed surprising similarities to human PSCs. NWR iPSCs exhibit a broad differentiation potency into the three germ layers and trophoblast, and acquire a naïve-like state of pluripotency, which is pivotal to differentiate PSCs into primordial germ cells (PGCs). Naïve culturing conditions induced a similar expression profile of pluripotency related genes in NWR iPSCs and human ESCs. Furthermore, naïve-like NWR iPSCs displayed increased expression of naïve and PGC marker genes, and a higher integration propensity into developing mouse embryos. As the conversion process was aided by ectopic BCL2 expression, and we observed integration of reprogramming factors, the NWR iPSCs presented here are unsuitable for gamete production. However, the gained insights into the developmental potential of both primed and naïve-like NWR iPSCs are fundamental for in future PGC-specification in order to rescue the species from extinction using cryopreserved somatic cells.

Cardiovascular and Metabolic Sciences Max Delbrück Center for Molecular Medicine in the Helmholtz Association 13125 Berlin Germany

Department of Developmental Genetics Max Planck Institute for Molecular Genetics 14195 Berlin Germany

Department of Stem Cell Biology and Medicine Graduate School of Medical Sciences Kyushu University Maidashi 3 1 1 Higashi ku Fukuoka 812 8582 Japan

Division of Drug Discovery and Safety Leiden Academic Centre for Drug Research Leiden University 2300 RA Leiden The Netherlands

Faculty of Veterinary Medicine Freie Universität Berlin 14163 Berlin Germany

FEMTransgenic Technologies Charité 13125 Berlin Germany

Fondazione Avantea 26100 Cremona Italy

Induced Pluripotent Stem Cell Core Facility Helmholtz Zentrum München 85764 Neuherberg Germany

Laboratory of Reproductive Technologies Avantea 26100 Cremona Italy

Leibniz Institute for Zoo and Wildlife Research 10315 Berlin Germany

Mobile DNA Max Delbrück Center for Molecular Medicine in the Helmholtz Association 13125 Berlin Germany

Technology Platform Pluripotent Stem Cells Max Delbrück Center for Molecular Medicine in the Helmholtz Association 13125 Berlin Germany

ZOO Dvůr Králové Štefánikova 1029 544 01 Dvůr Králové nad Labem Czech Republic

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In vitro fertilization program in white rhinoceros

Robust induction of primordial germ cells of white rhinoceros on the brink of extinction