Development of blastocyst complementation technology without contributions to gametes and the brain

• Published in: Experimental Animals Vol. 68; no. 3; pp. 361 - 370
• Main Authors: Hashimoto, Haruo, Eto, Tomoo, Yamamoto, Masafumi, Yagoto, Mika, Goto, Motohito, Kagawa, Takahiro, Kojima, Keisuke, Kawai, Kenji, Akimoto, Toshio, Takahashi, Ri-ichi
• Format: Journal Article
• Language: English
• Published: Japan Japanese Association for Laboratory Animal Science 2019; Japan Science and Technology Agency
• Subjects: Animals; blastocyst complementation; Blastocysts; Brain; Chimeras; Complementation; Embryo cells; Embryos; Gametes; Genes; Intelligence; Mice; Organs; Original; Otx2; Otx2 protein; Pluripotency; Prdm14; Reproductive organs; Stem cell transplantation; Stem cells; Technology; blastocyst complementation; Otx2; Prdm14
• ISSN: 1341-1357; 1881-7122
• DOI: 10.1538/expanim.18-0173

In Japan, it is possible to generate chimeric animals from specified embryos by combining animal blastocysts with human pluripotent stem (PS) cells (animal-human PS chimera). However, the production of animal-human PS chimeras has been restricted because of ethical concerns, such as the development of human-like intelligence and formation of humanized gametes in the animals, owing to the contributions of human PS cells to the brain and reproductive organs. To solve these problems, we established a novel blastocyst complementation technology that does not contribute to the gametes or the brain. First, we established GFP-expressing mouse embryonic stem cells (G-mESCs) in which the Prdm14 and Otx2 genes were knocked out and generated chimeric mice by injecting them into PDX-1-deficient blastocysts. The results showed that the G-mESCs did not contribute to the formation of gametes and the brain. Therefore, in the PDX-1-deficient mice complemented by G-mESCs without the Prdm14 and Otx2 genes, the germline was not transmitted to the next generations. This approach could address concerns regarding the development of both human gametes and a human-like brain upon mouse blastocyst complementation using human stem cells.