Pluripotent stem cells can be induced from somatic cells by the cellular intrinsic factors, including oocyte components or transcription factors1-4. Previously, we have demonstrated that mouse pluripotent stem cells can be induced from mouse somatic cells by small molecule-based external stimulation, providing a fundamentally different approach for inducing pluripotency5-8. Here we demonstrated that human pluripotent stem cells can be induced by small molecules from human somatic cells. This protocol describes the detailed procedures for the induction of human chemically induced pluripotent stem cells (hCiPSCs) from human somatic cells. In this method, the freshly isolated human embryonic fibroblasts (HEFs), human adipose derived mesenchymal stromal cells (hADSCs), and human adult skin fibroblasts are used for the induction of hCiPSCs. Four stages are used for the reprogramming process and the detailed induction medium for each stage is provided. In addition, we provide a troubleshooting list for the induction process.
We recently established a highly efficient and rapid chemical reprogramming system for generation of human pluripotent stem cells (hCiPSCs) (Liuyang et al., 2023). Here, we describe steps from isolation and culture of somatic cells to the derivation of hCiPSCs cell lines. In this method, human adipose derived stromal cells (hADSCs), and human adult skin fibroblasts (hASFs) are used as starting cells, we detail the preparation of small molecules stock solution and each stage conditional mediums, followed by step-by-step procedures to induction of hCiPSCs from somatic cells.
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