Abstract
Induced pluripotent stem cells (iPSCs) have considerably impacted human developmental biology and regenerative medicine, notably because they circumvent the use of cells of embryonic origin and offer the potential to generate patient-specific pluripotent stem cells. However, conventional reprogramming protocols produce developmentally advanced, or primed, human iPSCs (hiPSCs), restricting their use to post-implantation human development modeling. Hence, there is a need for hiPSCs resembling preimplantation naive epiblast. Here, we develop a method to generate naive hiPSCs directly from somatic cells, using OKMS overexpression and specific culture conditions, further enabling parallel generation of their isogenic primed counterparts. We benchmark naive hiPSCs against human preimplantation epiblast and reveal remarkable concordance in their transcriptome, dependency on mitochondrial respiration and X-chromosome status. Collectively, our results are essential for the understanding of pluripotency regulation throughout preimplantation development and generate new opportunities for disease modeling and regenerative medicine.
Highlights
Induced pluripotent stem cells have considerably impacted human developmental biology and regenerative medicine, notably because they circumvent the use of cells of embryonic origin and offer the potential to generate patient-specific pluripotent stem cells
We present a protocol enabling the parallel derivation of isogenic human induced primed and naive pluripotent stem cells. hiNPSCs are reprogrammed using T2iLGö7,19 or RSeT. hiNPSCs are benchmarked against the human preimplantation epiblast, the gold standard of human naive pluripotency, at the transcriptomic, metabolic and epigenetic levels
We aimed to develop a direct reprogramming method to simultaneously generate isogenic naive and primed human Pluripotent stem cells (PSCs)
Summary
Induced pluripotent stem cells (iPSCs) have considerably impacted human developmental biology and regenerative medicine, notably because they circumvent the use of cells of embryonic origin and offer the potential to generate patient-specific pluripotent stem cells. Significant progresses have been made to characterize the molecular signature of human preimplantation epiblast cells[10,11,12,13,14,15], establishing guidelines to assess human naive pluripotency[16] Those studies showed that two media supported naive pluripotent stem cells converted from primed cells or derived directly from human embryos, demonstrating hallmarks of human epiblast cells: 5i/L/AF8,17,18 and T2iLGö7,15,19,20. We present a protocol enabling the parallel derivation of isogenic human induced primed (hiPSCs) and naive (hiNPSCs) pluripotent stem cells. Direct somatic cell reprogramming to human naive pluripotency complements the array of assays enabling in-depth analysis of human pluripotency
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