Rspo1激活Lgr5+肝干细胞增强规模化培养肝类器官内细胞更新

肝类器官是体外培养具有肝功能的三维细胞聚集体,其培养技术的发展为肝脏研究领域提供了体外肝细胞持续培养的方法。实现肝类器官的规模化培养将极大推动肝细胞治疗的临床应用。研究表明，Wnt通路激动剂Rspo1蛋白不仅能在肝脏受损时激活肝内Lgr5+干细胞增殖，同时在体外培养中促进肝Lgr5+干细胞分裂生长形成类器官，展现出良好的细胞更新能力。在本研究中，将使用去细胞化肝脏胞外基质图案微阵列技术实现原代肝细胞、间充质干细胞和内皮细胞形成肝类器官的规模化培养，并探讨Rspo1蛋白能否诱导受损的类器官内Lgr5+肝干细胞增殖完成细胞自我更新，实现肝类器官长期培养。最后将规模化培养、具有更新能力的肝类器官用于生物人工肝（BAL）临床前治疗研究，评估其肝功能支持效果，促进临床转化。

Liver organoids are three-dimensional cell aggregates with liver function in vitro. The development of its culture technology provides a method for the continuous culture of hepatocytes in vitro for liver research. The realization of large-scale culture of liver organoids will greatly promote the clinical application of hepatocyte therapy. Studies have shown that the Wnt pathway agonist Rspo1 protein not only can activate Lgr5 + stem cell proliferation in the injured liver, but also promote liver Lgr5 + stem cells to divide and grow into organoids in vitro, showing good cell regeneration ability. In this study, decellularized liver ECM pattern microarray technology will be used to achieve large-scale culture of liver organoids formed via primary hepatocytes, mesenchymal stem cells and endothelial cells, and whether Rspo1 protein can induce injured organoids Lgr5 + liver stem cells proliferate to complete cell self-renewal and achieve long-term culture of liver organoids. Finally, large-scale cultured and renewable liver organoids were used for preclinical treatment of bio-artificial liver (BAL) to evaluate the effect of liver function support and promote clinical transformation.