Department of General Surgery, Tongji Hospital, School of Medicine, Tongji University, Shanghai, 200065, China.
Department of Ultrasonography, Tongji Hospital, School of Medicine, Tongji University, Shanghai, 200065, China.
Int J Med Sci. 2024 Jan 12;21(3):508-518. doi: 10.7150/ijms.83973. eCollection 2024.
This study aimed to explore the role of connexin 32 (Cx32) in the directional differentiation of induced pluripotent stem cells (iPSCs) into hepatocytes. Urine-derived epithelial cells were collected from the fresh urine of a healthy donor and transducted with reprogramming plasmid mixture to generate iPSCs. The iPSCs were then directionally differentiated into hepatocytes. During the differentiation, the upregulated and downregulated groups were treated with vitamin K2 (VK2) and 2-aminoethoxyboronate diphenylester (2-APB) to increase and inhibit Cx32 expression, respectively. The control group was not treated with the regulatory factor. Expression of Cx32 and hepatocyte-specific markers, including AFP, hepatocyte nuclear factor 4α (HNF-4α), albumin (ALB) and cytokeratin 18 (CK18) were detected. It indicated that Cx32 expression was not observed in iPSCs, but gradually increased during the process of hepatic differentiation from iPSCs. Upregulation of Cx32 expression by VK2 treatment promoted hepatocyte maturation and enhanced the expression of the aforementioned hepatic specific markers, whereas downregulation of Cx32 expression by 2-APB treatment had the opposite effects. In conclusion, urine-derived iPSCs could be directionally differentiated into hepatocytes. Up-regulation of Cx32 improves the efficiency and maturity of differentiation of iPSCs into hepatocytes, and Cx32 may be a promoting factor during the process of hepatic differentiation from iPSCs.
本研究旨在探讨连接蛋白 32（Cx32）在诱导多能干细胞（iPSCs）定向分化为肝细胞中的作用。从健康供体的新鲜尿液中收集尿上皮细胞，并转导重编程质粒混合物以生成 iPSCs。然后将 iPSCs 定向分化为肝细胞。在分化过程中，上调组和下调组分别用维生素 K2（VK2）和 2-氨基乙氧基硼酸盐二苯酯（2-APB）处理，以增加和抑制 Cx32 表达。对照组未用调节因子处理。检测 Cx32 和肝细胞特异性标志物的表达，包括 AFP、肝细胞核因子 4α（HNF-4α）、白蛋白（ALB）和细胞角蛋白 18（CK18）。结果表明，iPSCs 中未观察到 Cx32 表达，但在 iPSCs 向肝细胞分化的过程中逐渐增加。VK2 处理上调 Cx32 表达可促进肝细胞成熟，并增强上述肝特异性标志物的表达，而 2-APB 处理下调 Cx32 表达则产生相反的效果。总之，尿源 iPSCs 可定向分化为肝细胞。Cx32 的上调可提高 iPSCs 向肝细胞分化的效率和成熟度，Cx32 可能是 iPSCs 向肝细胞分化过程中的促进因子。
