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KMID : 0578320230460040209
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Molecules and Cells
2023 Volume.46 No. 4 p.209 ~ p.218
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Homogeneity of XEN Cells Is Critical for Generation of Chemically Induced Pluripotent Stem Cells
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Jeong Da-Hee
Lee Yu-Kyeong
Lee Seung-Won
Yanshuang Mu
Lee Min-Seong
Choi Na-Young
Guangming Wu
Hans R. Scholer
Ko Ki-Narm
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Abstract
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In induced pluripotent stem cells (iPSCs), pluripotency is induced artificially by introducing the transcription factors Oct4, Sox2, Klf4, and c-Myc. When a transgene is introduced using a viral vector, the transgene may be integrated into the host genome and cause a mutation and cancer. No integration occurs when an episomal vector is used, but this method has a limitation in that remnants of the virus or vector remain in the cell, which limits the use of such iPSCs in therapeutic applications. Chemical reprogramming, which relies on treatment with small-molecule compounds to induce pluripotency, can overcome this problem. In this method, reprogramming is induced according to the gene expression pattern of extra-embryonic endoderm (XEN) cells, which are used as an intermediate stage in pluripotency induction. Therefore, iPSCs can be induced only from established XEN cells. We induced XEN cells using small molecules that modulate a signaling pathway and affect epigenetic modifications, and devised a culture method in which can be produced homogeneous XEN cells. At least 4 passages were required to establish morphologically homogeneous chemically induced XEN (CiXEN) cells, whose properties were similar to those of XEN cells, as revealed through cellular and molecular characterization. Chemically iPSCs derived from CiXEN cells showed characteristics similar to those of mouse embryonic stem cells. Our results show that the homogeneity of CiXEN cells is critical for the efficient induction of pluripotency by chemicals.
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KEYWORD
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chemical reprogramming, chemically induced pluripotent stem cells, extra-embryonic endoderm, extra-embryonic endoderm cells, reprogramming
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