African Pygmy Mouse iPSCs as a Model for In Vitro Embryogenesis, Interspecies Chimerism, and Blastocyst Complementation

Somatic cell reprogramming into induced pluripotent stem cells (iPSCs) was achieved in various mammals, however assessing iPSC contribution to full-term chimeras beyond laboratory rodents remains challenging. Here, we demonstrate induction of pluripotency in male and female fibroblasts from the African Pygmy Mouse (APM), one of the smallest mammals. Using transcription factors and small molecules, we derived expandable APM-iPSCs that express pluripotency markers, differentiate into various cell types in vitro, and form gastruloids. Injection of APM-iPSCs into house mouse blastocysts generated full-term interspecies chimeras demonstrating extensive contribution to various tissues, including testicular germ cells. Notably, high APM contribution in organs such as heart and testes correlated with reduced organ size compared with mouse organs. Lastly, injection of APM-iPSCs into mouse blastocysts carrying a Pax7 ablation system enabled substantial production of APM muscle stem cells in chimeras. Collectively, this study establishes APM-iPSCs as a developmental model for pluripotency, differentiation and interspecies chimerism. Overall design: Mouse-APM interspecies chimeras were produced through blastocyst injection into the Pax7-Cre/ERT2; Rosa26-loxP-STOP-loxP-Diphtheria toxin A (DTA) (B6.Cg-Pax7tm1(cre/ERT2)Gaka/J; B6.129P2-Gt(ROSA)26Sortm1(DTA)Lky/J) host, followed by tamoxifen-induced ablation of Pax7 expressing host cells.