Animal Reproduction Institute, State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi University, Nanning, P.R. China.
International Zhuang Medical Hospital Affiliated to Guangxi University Chinese Medicine, Nanning, P.R. China.
Cell Reprogram. 2021 Jun;23(3):158-167. doi: 10.1089/cell.2020.0094. Epub 2021 May 5.
Past researches have shown that pluripotency maintenance of naive and primed-state pluripotent stem cells (PSCs) depends on different signaling pathways, and naive-state PSCs possess the ability to produce chimeras when they are introduced into a blastocyst. Considering porcine is an attractive model for preclinical studies, many researches about pig induced pluripotent stem cells (piPSCs) have been reported. Some cytokines and small molecule compounds could transform primed piPSCs into naive state. However, there are no suitable culture conditions for generation of naive-state piPSCs with high efficiency; other small molecule compounds need further exploration. In this study, we investigated whether p38 MAPK and JNK signal pathway inhibitor SB203580 and SP600125 could be of benefit for acquiring naive-state piPSCs. By comparing reprogramming efficiencies under conditions of different donor cells and culture environment, we found that porcine bone marrow mesenchymal stem cells (PBMSCs) have higher efficiency on piPSC induction, and the culture condition of CHIR99021+PD0325901(2i)+Lif+bFGF is more suitable for subculturing of piPSCs. Our results also indicate that SB203580 and SP600125 could promote reprogramming of PBMSCs into naive-like state piPSCs. These results provide guidance for choosing donor cells, culture conditions, and research of different state iPSCs during the process of reprogramming pig somatic cells.
过去的研究表明，原始态和起始态多能干细胞（PSCs）的多能性维持依赖于不同的信号通路，而且原始态 PSCs 具有在囊胚中产生嵌合体的能力。考虑到猪是临床前研究的有吸引力的模型，已经有许多关于猪诱导多能干细胞（piPSCs）的研究报告。一些细胞因子和小分子化合物可以将起始态 piPSCs 转化为原始态。然而，目前还没有高效生成原始态 piPSCs 的合适培养条件；其他小分子化合物需要进一步探索。在这项研究中，我们研究了 p38 MAPK 和 JNK 信号通路抑制剂 SB203580 和 SP600125 是否有利于获得原始态 piPSCs。通过比较不同供体细胞和培养环境下的重编程效率，我们发现猪骨髓间充质干细胞（PBMSCs）在诱导 piPSC 方面具有更高的效率，并且 CHIR99021+PD0325901（2i）+Lif+bFGF 的培养条件更适合 piPSC 的传代培养。我们的结果还表明，SB203580 和 SP600125 可以促进 PBMSCs 向原始样态 piPSCs 的重编程。这些结果为选择供体细胞、培养条件以及在猪体细胞重编程过程中研究不同状态的 iPSCs 提供了指导。
