CHIR99021 and Brdu Are Critical in Chicken iPSC Reprogramming via Small-Molecule Screening

文献类型： 外文期刊

第一作者： Jin, Kai

作者： Jin, Kai;Zhou, Jing;Wu, Gaoyuan;Li, Zeyu;Zhu, Xilin;Liang, Youchen;Li, Tingting;Chen, Guohong;Zuo, Qisheng;Niu, Yingjie;Jin, Kai;Zhou, Jing;Wu, Gaoyuan;Li, Zeyu;Zhu, Xilin;Liang, Youchen;Li, Tingting;Chen, Guohong;Zuo, Qisheng;Niu, Yingjie;Jin, Kai;Zhou, Jing;Wu, Gaoyuan;Li, Zeyu;Zhu, Xilin;Liang, Youchen;Li, Tingting;Chen, Guohong;Zuo, Qisheng;Niu, Yingjie;Jin, Kai;Song, Jiuzhou;Han, Wei

作者机构：

关键词： CHIR99021; BrdU; chicken iPSC; small molecule; reprograming

期刊名称：GENES （ 影响因子：2.8； 五年影响因子：3.2 ）

ISSN：

年卷期： 2024 年 15 卷 9 期

页码：

收录情况： SCI

摘要： Background/Objectives: Induced pluripotent stem cells (iPSCs) reprogrammed from somatic cells into cells with most of the ESC (embryonic stem cell) characteristics show promise toward solving ethical problems currently facing stem cell research and eventually yield clinical grade pluripotent stem cells for therapies and regenerative medicine. In recent years, an increasing body of research suggests that the chemical induction of pluripotency (CIP) method can yield iPSCs in vitro, yet its application in avian species remains unreported. Methods: Herein, we successfully obtained stably growing chicken embryonic fibroblasts (CEFs) using the tissue block adherence method and employed 12 small-molecule compounds to induce chicken iPSC formation. Results: The final optimized iPSC induction system was bFGF (10 ng/mL), CHIR99021 (3 mu M), RepSox (5 mu M), DZNep (0.05 mu M), BrdU (10 mu M), BMP4 (10 ng/mL), vitamin C (50 mu g/mL), EPZ-5676 (5 mu M), and VPA (0.1 mM). Optimization of the induction system revealed that the highest number of clones was induced with 8 x 104 cells per well and at 1.5 times the original concentration. Upon characterization, these clones exhibited iPSC characteristics, leading to the development of a stable compound combination for iPSC generation in chickens. Concurrently, employing a deletion strategy to investigate the functionality of small-molecule compounds during induction, we identified CHIR99021 and BrdU as critical factors for inducing chicken iPSC formation. Conclusions: In conclusion, this study provides a reference method for utilizing small-molecule combinations in avian species to reprogram cells and establish a network of cell fate determination mechanisms.

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