Laboratory of Molecular Pharmacology, Department of Pharmacology, School of Pharmacy, Southwest Medical University, Luzhou, People's Republic of China.
Key Laboratory for Regenerative Medicine of the Ministry of Education of China, School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, People's Republic of China.
Stem Cells Transl Med. 2024 Feb 14;13(2):166-176. doi: 10.1093/stcltm/szad079.
Cholangiocytes form a complex 3D network of bile ducts in the liver and contribute to liver function. The damage or destruction of cholangiocytes can lead to biliary diseases, and the shortage of cholangiocytes remains an obstacle for drug development targeting biliary diseases. Valproic acid (VPA) is a potent activator of Notch signaling pathway that is essential for cholangiocyte differentiation. Here, we report a VPA-based approach for cholangiocyte differentiation of human pluripotent stem cells. VPA activated Notch2 expression and upregulated HES-1, HEY-1, and Sox9 gene expression in hESC-derived hepatoblast. After 7 days treatment, VPA promoted successful differentiation of hepatoblast into cholangiocytes expressing cholangiocyte marker genes (AE2, AQP1, CFTR) and proteins (CK19 and CK7). In addition, the differentiated cholangiocytes formed bile duct-like structures after implantation into the spleen of NOD/SCID mice. Our results suggested that VPA can promote hESC differentiation to cholangiocyte-like cells. The induced cholangiocytes may serve as a potential cell source for both in vitro modeling and regenerative therapy of cholangiopathies. The findings can also support further development of small-molecule based differentiation protocols for cholangiocyte production.
胆管细胞在肝脏中形成复杂的三维胆管网络，并有助于肝脏功能。胆管细胞的损伤或破坏可导致胆管疾病，而胆管细胞的短缺仍然是针对胆管疾病的药物开发的障碍。丙戊酸（VPA）是 Notch 信号通路的有效激活剂，对胆管细胞分化至关重要。在这里，我们报告了一种基于 VPA 的人多能干细胞胆管细胞分化方法。VPA 激活了 hESC 来源的肝母细胞中的 Notch2 表达，并上调了 HES-1、HEY-1 和 Sox9 基因的表达。经过 7 天的处理，VPA 促进了肝母细胞成功分化为表达胆管细胞标记基因（AE2、AQP1、CFTR）和蛋白（CK19 和 CK7）的胆管细胞。此外，分化的胆管细胞在植入 NOD/SCID 小鼠的脾脏后形成胆管样结构。我们的结果表明，VPA 可以促进 hESC 分化为胆管样细胞。诱导的胆管细胞可作为胆管病体外建模和再生治疗的潜在细胞来源。这一发现也可以支持进一步开发基于小分子的胆管细胞生产分化方案。
