College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology, Northwest A&F University, Yangling, China.
Cell Prolif. 2021 Jan;54(1):e12932. doi: 10.1111/cpr.12932. Epub 2020 Oct 26.
The establishment of porcine pluripotent stem cells (pPSCs) is still a critical topic. However, all pPSCs were failed to contribute to efficient chimeric pig and were extremely sensitive to changes of culture conditions. This study aimed to investigate the role of BCL2 in pPSCs and further explain the mechanism.
Porcine BCL2 gene was cloned and overexpressed in porcine induce pluripotent stem cells (piPSCs). Digital RNA-seq was performed to explain the mechanism of anti-apoptosis. Finally, the cells carrying BCL2 were injected into mouse early embryo to evaluate its chimeric ability.
Here, we found that overexpression of porcine BCL2 gene significantly improved the survivability of piPSCs and the efficiency of embryonic chimerism, and did not wreck the pluripotency of piPSCs. Furthermore, the Digital RNA-seq analysis revealed that BCL2, as a downstream gene of the PI3K signal pathway, enhanced the expression of PI3K signal pathway receptors, such as FGFR2, and further promoted oxidoreductases activity and lipid metabolism, thus maintaining the survival and pluripotency of piPSCs.
Our data not only suggested that porcine BCL2 gene could enhance the survivability and chimeric ability of pPSCs, but also explained the positive feedback mechanism in this process, providing strong support for the chimeric experiment of pPSCs.
猪多能干细胞（pPSCs）的建立仍是一个关键问题。然而，所有的 pPSCs 都未能有效地促成嵌合猪的产生，并且对培养条件的变化极其敏感。本研究旨在探讨 BCL2 在 pPSCs 中的作用，并进一步解释其机制。
克隆猪 BCL2 基因并在猪诱导多能干细胞（piPSCs）中过表达。进行数字 RNA-seq 以解释抗凋亡的机制。最后，将携带 BCL2 的细胞注入小鼠早期胚胎中，以评估其嵌合能力。
我们发现，猪 BCL2 基因的过表达显著提高了 piPSCs 的存活率和胚胎嵌合效率，且并未破坏 piPSCs 的多能性。此外，数字 RNA-seq 分析表明，BCL2 作为 PI3K 信号通路的下游基因，增强了 PI3K 信号通路受体，如 FGFR2 的表达，进而促进了氧化还原酶活性和脂质代谢，从而维持 piPSCs 的存活和多能性。
我们的数据不仅表明猪 BCL2 基因可以提高 pPSCs 的存活率和嵌合能力，还解释了这一过程中的正反馈机制，为 pPSCs 的嵌合实验提供了有力支持。
