2023.12.11文献分享【Rolling back human pluripotent stem cells to an eight-cell embryo-like stage】

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最新推荐文章于 2025-05-03 09:08:56 发布

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【Title】Rolling back human pluripotent stem cells to an eight-cell embryo-like stage

【Publication Book】Nature

【Publication Time】2023 March

【Abstract】Here, author described a transgene-free, rapid and controllable method for producing 8C-like cells (8CLCs) from human pluripotent stem cells.

【Background】

Human PSCs can be maintained in naive or primed states, which reflect the preimplantation and postimplantation ICM, respectively.

Mouse naive PSC cultures contain a small proportion (approximately 0.5%) of cells sharing features of the two-cell (2C) embryonic stage, a time when mouse zygotic genome activation (ZGA) occurs and totipotency is achieved. These cells are termed 2C-like cells (2CLCs), and their proportion can be enhanced through manipulation.

Despite advancements achieved in methods using rodent models, the generation of 8CLCs that transcriptionally and epigenetically match the human 8C embryo, and evaluation of their functional properties, have not yet been described

【Result】

1.从人类 PSC 中生成 8CLC

1.1 In the first round, combining the MEK inhibitor PD0325901, the tankyrase inhibitor IWR1 and human leukaemia inhibitory factor (LIF) in a chemically defined medium (N2B27, activin A and vitamin C14) on feeder cells produced PSC colonies with a naive PSC-like colony morphology and the highest levels of naive pluripotency gene expression, as assessed by quantitative PCR with reverse transcription (RT–qPCR)

PD+IWR1+hLIF

1.2 In round two, author tested 14 additional compounds added to this basal medium.DZNep+TSA+PD+IWR1+hLIF substantially increased the expression of naive pluripotency genes

DZNep+TSA+PD+IWR1+hLIF (4 chemicals+LIF)——4CL

Naive PSC marked: OCT4 NANOG DPPA3 DPPA5 MAEL KLF4 KLF17

1.3 Author also noted that removing vitamin C reduced the expression of naive pluripotency genes in the 4CL-mediated conversion process. Moreover, TSA could be effectively substituted by other HDAC inhibitors such as valproic acid o