Deng Yanfei, Lao Yanping, Ruan Qiuyan, Zhang Jun, Luo Chan, Shi Deshun, Lu Fenghua
Animal Reproduction Institute, State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi University, Guangxi, China.
Cell Reprogram. 2020 Aug;22(4):217-225. doi: 10.1089/cell.2020.0027. Epub 2020 Jul 14.
Wnt/β-Catenin signaling pathway plays an important role in maintaining self-renewal and pluripotency of human and mouse embryonic stem cells (ESCs). Activation of Wnt/β-Catenin signaling pathway by glycogen synthase kinase-3 (GSK3) inhibitor, the Wnt signaling agonist, could maintain the pluripotency of human and mouse ESCs in the presence of serum. However, the role of signaling pathway in the derivation of buffalo ESCs remains unclear. In this study, we used GSK3 inhibitors (6-bromoindirubin-3'-oxime [BIO] and CHIR99021) and investigated the effect of Wnt/β-Catenin activation on colony formation, proliferation, self-renewal, and pluripotency of Chinese swamp buffalo (buffalo) embryonic stem cell-like cells (ES-like cells), which were isolated from blastocysts. The results showed that buffalo ES-like cells displayed typical morphological characteristics of pluripotent stem cells: positive for alkaline phosphatase staining, expression of pluripotent markers, including , , SSEA-1, SSEA-4, , , , and the proliferative markers, and . Furthermore, activation of Wnt/β-Catenin signaling pathway by GSK3 inhibitors could promote colony formation and proliferation of buffalo ES-like cells and maintain their undifferentiated state, and upregulate the expression levels of pluripotent-related genes and proliferation-related genes. These results indicated that Wnt/β-Catenin signaling pathway plays an important role in the derivation and pluripotency of buffalo ES-like cells.
Wnt/β-连环蛋白信号通路在维持人类和小鼠胚胎干细胞（ESC）的自我更新和多能性方面发挥着重要作用。糖原合酶激酶-3（GSK3）抑制剂（Wnt信号激动剂）激活Wnt/β-连环蛋白信号通路，可在有血清存在的情况下维持人类和小鼠ESC的多能性。然而，该信号通路在水牛ESC的分离过程中的作用仍不清楚。在本研究中，我们使用了GSK3抑制剂（6-溴靛玉红-3'-肟[BIO]和CHIR99021），并研究了Wnt/β-连环蛋白激活对从囊胚中分离出的中国沼泽水牛（水牛）胚胎干细胞样细胞（ES样细胞）的集落形成、增殖、自我更新和多能性的影响。结果表明，水牛ES样细胞表现出多能干细胞的典型形态特征：碱性磷酸酶染色呈阳性，多能性标志物表达阳性，包括 、 、阶段特异性胚胎抗原-1（SSEA-1）、阶段特异性胚胎抗原-4（SSEA-4）、 、 、 ，以及增殖标志物 和 。此外，GSK3抑制剂激活Wnt/β-连环蛋白信号通路可促进水牛ES样细胞的集落形成和增殖，并维持其未分化状态，上调多能相关基因和增殖相关基因的表达水平。这些结果表明，Wnt/β-连环蛋白信号通路在水牛ES样细胞的分离和多能性方面发挥着重要作用。
