Rat embryonic stem cells produce fertile offspring through tetraploid complementation

Abstract

Pluripotency of embryonic stem cells (ESCs) can be functionally assessed according to the developmental potency. Tetraploid complementation, through which an entire organism is produced from the pluripotent donor cells, is taken as the most stringent test for pluripotency. It remains unclear whether ESCs of other species besides mice can pass this test. Here we show that the rat ESCs derived under 2i (two small molecule inhibitors) conditions at very early passages are able to produce fertile offspring by tetraploid complementation. However, they lose this capacity rapidly during culture due to a nearly complete loss of genomic imprinting. Our findings support that the naïve ground state pluripotency can be captured in rat ESCs but also point to the species-specific differences in its regulation and maintenance, which have implications for the derivation and application of naïve pluripotent stem cells in other species including human.

Blastocyst complementation using Prdm14-deficient rats enables efficient germline transmission and generation of functional mouse spermatids in rats

“…However, rat PSCs are less robust than mouse PSCs, partly because of the difficulties of handling and clonal selection after genetic manipulation 20 , 21 . Besides, rat PSCs exhibit rapid loss of genomic imprints in culture, which affects their developmental potential, as judged by tetraploid complementation experiments designed to produce an entirely PSC-derived animal 22 , 23 . Here, we show that the use of rat blastocyst from Prdm14 KO rats serve as excellent hosts for the germline transmission of donor cells.…”

Silencing of retrotransposon-derived imprinted gene RTL1 is the main cause for postimplantational failures in mammalian cloning

“…Importantly, when reanalyzing the SC datasets from previous studies ( 53 – 55 ) we noticed that the expression of RTL1 could be a marker to evaluate developmental potential of SCs in mammals. We found that 4N “on” mouse iPSCs (competent for tetraploid complementation) showed higher RTL1 expression than 4N “off” mouse iPSCs (incompetent for tetraploid complementation) ( Fig.…”

Domesticated cynomolgus monkey embryonic stem cells allow the generation of neonatal interspecies chimeric pigs

“…Interspecies chimerism has been most successful in rodents, and these studies typically achieve a high proportion of chimerism and generate xenogeneic organs (Kobayashi et al, 2010;Isotani et al, 2011). The pluripotency levels of rodent PSCs are the highest known, as they are genuine naïve cells, and they can produce live offspring by tetraploid complementation, which is the most stringent (Zhao et al, 2009;Li et al, 2017). Recent studies on xenogeneic chimerism in mice and rats showed that the pluripotency of rat ESCs affected kidney organ reconstruction in mice (Goto et al, 2019), demonstrating that the pluripotency of the donor cells influenced xenogeneic chimerism.…”