Here, we show that rhinoceros oocytes can be repeatedly recovered from live SWR females by transrectal ovum pick-up (OPU), matured in vitro, fertilized by intracytoplasmic sperm injection (ICSI) and, for the first time, develop to the blastocyst stage in vitro. Surplus embryos can be cryopreserved for later transfer or processed as sources of stem cell. Reproduction through assisted medical procedures requires safely harvesting oocytes and spermatozoa, achieving in vitro maturation of oocytes, in vitro fertilization, stimulating the resulting zygote to grow to the blastocyst stage, and safely and efficiently transferring the blastocyst to the uterus of a synchronous surrogate mother. They are large, somewhat intractable animals, bigger than horse and difficult to manipulate, and ART has never been completely successful in this species 1. Using this technology, the NWR faces some unique challenges there are only two surviving NWRs and both are infertile. The rhinoceros and horse share a common ancestor 6, therefore, assisted reproduction techniques (ART) developed in equines 7, 8 can potentially be translated to rhinoceros species, including the Southern White Rhinoceros (SWR, Ceratotherium simum simum), a closely related sub-species now not at risk of extinction 9. A holistic strategy combining all three steps would help to maximize genetic diversity in the NWR by providing cryo-banked spermatozoa and somatic cells of several NWR individuals 5. ES cells are the best source for producing primordial germ cells and also represent the “gold standard” for assessing artificial gametes derived from induced pluripotent stem cells (iPSCs) 4. Beyond applying this classical ART approach, it would be critical to generate artificial gametes. Our results suggest that ART might offer an option for rescuing genes from the NWR, an essential first step in saving this nearly extinct rhinoceros sub-species. There has been no report of the production of rhinoceros embryo from fertilization to the pre-implantation stage 2, 3. To date only one study related to ovum pick up and embryos production in rhinoceros has been reported 1. Conventional approaches toward establishing a self-sustaining NWR population over the last two decades have been repeatedly unsuccessful.