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Gene expression in primed and chemically reset (cR) naïve human iPSCs, and naïve-converted cells after co-culture with mouse embryonic stem cells

ABSTRACT: Gene expression in primed and chemically reset (cR) naïve human iPSCs, and naïve-converted cells after co-culture with mouse embryonic stem cells

PROVIDER: PRJNA893005 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR22007816_1.fastq.gz	Fastqsanger.gz
	SRR22007816_2.fastq.gz	Fastqsanger.gz
	SRR22007817_1.fastq.gz	Fastqsanger.gz
	SRR22007817_2.fastq.gz	Fastqsanger.gz
	SRR22007818_1.fastq.gz	Fastqsanger.gz

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Gene expression in primed and chemically reset (cR) naïve human iPSCs, and naïve-converted cells after co-culture with mouse embryonic stem cells

Project description:We compared the global transcriptome between mESC co-culture-mediated naïve-like hiPSCs and cR cells together with their parental primed cells.

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Project description:Functional-assay limitations are an emerging issue in characterizing human pluripotent stem cells (hPSCs). With rodent PSCs, chimera formation, using pre-implantation embryos, is the gold-standard assay of pluripotency. In hPSCs, this can only be monitored via teratoma formation or in vitro differentiation, as ethical concerns preclude generation of human-animal chimera. To circumvent this issue, we established a functional assay utilizing interspecific blastocyst injection and in vitro culture (interspecies in vitro chimera assay). The assay uses mouse pre-implantation embryos and human PSCs to make interspecies chimeras cultured in vitro to the early egg cylinder stage. When hiPSCs, both conventional and naive type, which called M-bM-^@M-^\reset cellM-bM-^@M-^], were injected into mouse embryos and cultured. The cells were never integrated into the epiblast of egg cylinder stage-embryo. These results suggest that hPSCs, including naM-CM-/ve type, are unable to form chimera with mouse embryo. Reset cells were converted from conventional human iPSC line PB004, and then compared their gene expression profile with or without transgene overexpression induced by doxycyclin treatment.

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Project description:Functional-assay limitations are an emerging issue in characterizing human pluripotent stem cells (hPSCs). With rodent PSCs, chimera formation, using pre-implantation embryos, is the gold-standard assay of pluripotency. In hPSCs, this can only be monitored via teratoma formation or in vitro differentiation, as ethical concerns preclude generation of human-animal chimera. To circumvent this issue, we established a functional assay utilizing interspecific blastocyst injection and in vitro culture (interspecies in vitro chimera assay). The assay uses mouse pre-implantation embryos and human PSCs to make interspecies chimeras cultured in vitro to the early egg cylinder stage. When hiPSCs, both conventional and naive type, which called “reset cell”, were injected into mouse embryos and cultured. The cells were never integrated into the epiblast of egg cylinder stage-embryo. These results suggest that hPSCs, including naïve type, are unable to form chimera with mouse embryo.

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