人ESC来源的共表达凝血因子Ⅷ、Ⅸ的功能性肝细胞血友病动物模型替代治疗研究

The research of cell replacement therapy for hemophilia has broad prospects, use of embryonic stem cells (ESC) derived hepatocytes coexpressing coagulation factor VIII&IX（F-VIII&IX）as cell source attracts the public eye.But the current domestic and international reports on human ESC-derived hepatocytes replacement therapy is still very limited. Our previous studies have established an in vitro multi-factor differentiation protocol for mouse ESC-derived hepatocytes coexpressing F-VIII&IX, and the feasibility of in vivo transplantation experiments have also been confirmed. On this basis,we initiate the research of human ES-derived hepatocytes.As a result, the project intends to carry out the following studies: ① Establishment of efficient in vitro differentiation protocol of human ESC-derived functional hepatocytes coexpressing F-VIII&IX; ②In vivo transplantation of the human ESC-derived hepatocytes as well as the efficacy evaluation system will be studied to verify the efficacy and safety of stem cell replacement therapy for hemophilia. This study aimed to explore the important value of embryonic stem cell replacement therapy for hemophilia, and then lay a more solid experimental basis for preclinical studies.

表达凝血因子VIII、IX的胚胎干细胞（ESC）源性肝细胞移植替代治疗血友病具有广阔的研究前景，但目前国内外关于人ESC源性肝细胞血友病替代治疗研究的报道仍十分有限。我们的前期研究证明，多因子联合培养体系能在体外诱导小鼠胚胎干细胞分化为稳定表达凝血因子VIII、IX的功能性肝细胞，并进行了动物体内移植实验，将此培养体系用于人ESC肝向诱导分化也经初步验证可行。由此，本项目拟开展以下研究：①建立高效的体外诱导人ESC分化为稳定表达凝血因子VIII、IX的功能性肝细胞培养体系；②将人ESC源性肝细胞移植入血友病裸鼠模型体内，建立体内移植、替代疗效评价体系，验证干细胞替代疗法治疗血友病的有效性和安全性。本研究旨在探索人ESC源性肝细胞在血友病治疗中的重要价值，从而为血友病胚胎干细胞替代疗法的临床前研究奠定更坚实的实验基础。

表达凝血因子VIII、IX的胚胎干细胞（ESC）源性肝细胞移植替代治疗血友病具有广阔的研究前景，但目前国内外关于人ESC源性肝细胞血友病替代治疗研究的报道仍十分有限。我们的前期研究证明，多因子联合培养体系能在体外诱导小鼠胚胎干细胞分化为稳定表达凝血因子VIII、IX的功能性肝细胞，并进行了动物体内移植实验，将此培养体系用于人ESC肝向诱导分化也经初步验证可行。由此，本项目在前期基础上开展了以下研究：①建立了高效的体外诱导人ESC分化为稳定表达凝血因子VIII、IX的功能性肝细胞培养体系；②将人ESC源性肝细胞移植入血友病裸鼠模型体内，建立了体内移植、替代疗效评价体系，验证干细胞替代疗法治疗血友病的有效性和安全性。本研究旨在探索人ESC源性肝细胞在血友病治疗中的重要价值，从而为血友病胚胎干细胞替代疗法的临床前研究奠定更坚实的实验基础。