定向分化人间充质干细胞成为胆管细胞作为新型体外疾病模型的研究

Current treatment options remain largely ineffective, necessitating liver transplantation as the only definitive treatment in most patients with advanced disease. The development of effective pharmacological agents for bile duct disorders has been hampered by poor access to primary biliary tissue, difficulties in culturing primary cholangiocytes in vitro and inadequate disease models. We have established defined culture conditions to derive cholangiocyte-like cells from human mesenchymal stem cells, and developed an imaging platform to elucidate the short- and long-term in vivo fate of administered mesenchymal stem cells. This project first aims to improve the differentiation efficiency of mesenchymal stem cells into cholangiocyte-like cells, study their in vivo and in vitro function, and establish the in vitro disease models and drug-testing platform. The capacity of our platform for generating large amounts of normal and disease-specific functional cholangiocytes will have broad applications in tissue engineering, in disease modelling and for screening of therapeutic compounds for bile duct disorders.

目前胆道系统疾病尚无有效的治疗手段，对于晚期胆道系统疾病患者来说，肝脏移植往往是唯一的治疗手段。另一方面，针对胆道系统疾病的药物研发进展缓慢，其主要原因是临床上难以获得原位胆道组织、胆管细胞体外培养困难，以及缺乏合适的疾病模型。我们的前期研究发现间充质干细胞可以通过特定的体外培养基定向诱导分化为有功能的胆管样细胞，同时我们建立了体内追踪外源性间充质干细胞的活体成像平台。本研究将首先优化定向分化步骤，尝试将间充质干细胞高效地定向分化为有功能的胆管样细胞，再从体外、体内两个研究层面完整地研究从这种的胆管样细胞的功能，并尝试建立以此细胞为基础的体外疾病模型和药敏试验平台。本项目中成功获得的胆管样细胞将有助于针对胆道系统疾病的药物筛选，同时也可应用于胆管重建，具有广阔的应用前景。

本研究设计了定向分化步骤，该“两步分化法”不需要将干细胞和其他细胞共培养或基因重组，仅仅依靠特定培养条件诱导分化。成功将间充质干细胞高效地定向分化为有功能的胆管样细胞，第27天分化得到的胆管细胞高表达胆管标志物（CK7、CK19、TGR5、CFTR 和AQP1），低表达肝细胞标志物。本研究再从体外、体内两个研究层面完整地研究从这种的胆管样细胞的功能，本项目中成功获得的胆管样细胞将有助于针对胆道系统疾病的药物筛选，同时也可应用于胆管重建，具有广阔的应用前景。共发表相关论文两篇。