Abstract
SummaryPluripotent stem cells are defined by their capacity to differentiate into all three tissue layers that comprise the body. Chimera formation, generated by stem cell transplantation to the embryo, is a stringent assessment of stem cell pluripotency. However, the ability of human pluripotent stem cells (hPSCs) to form embryonic chimeras remains in question. Here we show using a stage-matching approach that human induced pluripotent stem cells (hiPSCs) and human embryonic stem cells (hESCs) have the capacity to participate in normal mouse development when transplanted into gastrula-stage embryos, providing in vivo functional validation of hPSC pluripotency. hiPSCs and hESCs form interspecies chimeras with high efficiency, colonize the embryo in a manner predicted from classical developmental fate mapping, and differentiate into each of the three primary tissue layers. This faithful recapitulation of tissue-specific fate post-transplantation underscores the functional potential of hPSCs and provides evidence that human-mouse interspecies developmental competency can occur.
Highlights
For mESCs, confirmation of stem cell pluripotency includes a demonstration of their ability to integrate into the preimplantation embryo and subsequently contribute to all the tissues of the developing mouse chimera (Bradley et al, 1984; Nagy et al, 1993)
We transplanted three human induced pluripotent stem cells (hiPSCs) and two human embryonic stem cells (hESCs) lines, each transfected with a fluorescent reporter gene, into early and late gastrula-stage mouse embryos at the primitive streak or distal tip of the epiblast (Figure 1A)
We found highly efficient interspecies chimera formation in all transplant sites ranging from 70% to 100% of transplanted embryos following in vitro culture (Figure 1B). (Embryos were obtained and cultured under University ethical review according to UK animal regulations; see Supplemental Experimental Procedures.) We transplanted mEpiSCs in a similar manner as a positive control for a putative interspecies barrier and saw similar incorporation
Summary
For mESCs, confirmation of stem cell pluripotency includes a demonstration of their ability to integrate into the preimplantation embryo and subsequently contribute to all the tissues of the developing mouse chimera (Bradley et al, 1984; Nagy et al, 1993). We transplanted three hiPSC and two hESC lines (together, hPSCs), each transfected with a fluorescent reporter gene, into early and late gastrula-stage mouse embryos at the primitive streak or distal tip of the epiblast (Figure 1A).
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