体外3D规模化扩增肝细胞的培养体系及自动化、智能化生物反应器的评估  

Assessment of in vitro 3D large-scale hepatocyte proliferation culture system and automated and intelligent bioreactor systems

作　　者：李柯[1,2] 张广浩 张丞[1,2] 杨嘉屹 吴昌哲[1,2,3] 霍小林 Li Ke;Zhang Guanghao;Zhang Cheng;Yang Jiayi;Wu Changzhe;Huo Xiaolin(Beijing Key Laboratory of Bioelectromagnetism,Institute of Electrical Engineering,Chinese Academy of Sciences,Beijing 100190,China;University of Chinese Academy of Sciences,Beijing 100049,China;ZhuJiang Hospital of Southern Medical University,Guangzhou 510280,Guangdong Province,China)

机构地区：[1]中国科学院电工研究所生物电磁学北京市重点实验室,北京市100190 [2]中国科学院大学,北京市100049 [3]南方医科大学珠江医院,广东省广州市510280

出　　处：《中国组织工程研究》2022年第19期3100-3107,共8页Chinese Journal of Tissue Engineering Research

基　　金：国家重点研发计划(2018YFC1106400,2018YFA0108200),项目参与人:吴昌哲;国家自然科学基金(61671428),项目负责人:吴昌哲。

摘　　要：背景:如何在体外获得足够数量和质量的肝细胞,是生物人工肝临床应用亟待解决的核心问题。目的:对肝细胞体外3D规模化扩增培养体系的主要研究内容进行综述,总结了当前主要的肝种子细胞来源、3D培养方法、生物反应器系统以及培养过程关键指标实时在线监测的进展和局限,并对自动化、智能化生物反应器系统进一步发展进行展望。方法:由第一作者检索Web of Science核心数据库、ScienceDirect、EI、PubMed以及中国知网数据库,英文检索词为:"hepatocyte,3D culture,proliferate,bioreactorsystem,monitor andcontrol",中文检索词为:"肝细胞、3D培养、增殖、生物反应器系统、监测和控制"。文献检索时间范围为2005年1月至2021年3月,文献的类型包括研究原著、综述和荟萃分析,最终筛选出93篇文献进行综述。结果与结论:(1)目前肝种子细胞来源、3D培养方式、生物反应器系统构建以及关键生化指标实时在线监测方法等方面的研究均取得了一定进展,为进一步构建肝细胞体外3D规模化培养体系奠定了坚实的基础。(2)人肝癌细胞系、永生化肝细胞、干细胞分化及其他来源的肝细胞均能在体外培养增殖并进行肝功能表达,其中干细胞分化和直接重编程得到的肝细胞样细胞具有成熟肝脏细胞类似的细胞形态、功能和基因表达谱特征,在未来有更大的应用前景。(3)3D悬浮培养在培养密度、传质效率方面表现优异,且具有易于放大、接种和取用方便等优点,是肝细胞体外3D培养方式的主要发展方向。(4)现有的生物反应器系统虽然能实现肝细胞体外培养扩增,但自动化程度低,且缺乏对关键生化指标进行实时在线检测方法;此外,当前这几方面的研究相互融合程度低,也制约了肝细胞体外3D规模化培养体系的进展。(5)未来的研究重点是结合不同来源的肝种子细胞的生长特性,选择适合培养方式,以建立BACKGROUND:How to obtain sufficient quantity and quality of hepatocytes in vitro is the core problem to be solved urgently in the clinical application of bioartificial liver.OBJECTIVE:To summarize the main research contents of the in vitro 3D large-scale expansion culture system of hepatocytes,summarize the current main sources of seed cells,3D culture methods,bioreactor system,and the progress and limitations of real-time online monitoring of key indicators of the culture process,and prospects for the further development of automated and intelligent bioreactor systems.METHODS:The first author searched the Web of Science core database,ScienceDirect,EI,PubMed,and CNKI databases.The English and Chinese search terms were"hepatocyte,3D culture,proliferate,bioreactor system,monitor and control".The literature search time range was from January 2005 to March 2021.The types of articles included original research works,reviews and meta-analysis.Totally,93 papers were selected for analysis.RESULTS AND CONCLUSION:(1)Researches on liver seed cell sources,3D culture methods,bioreactor system construction,and real-time online monitoring methods for key biochemical indicators have all made progress,laying a solid foundation for the further construction of a large-scale in vitro 3D hepatocyte culture system.(2)Human hepatocarcinoma cell lines,immortalized hepatocytes,stem cell differentiation and other hepatocytes can be cultured and proliferated in vitro and express liver function.Among them,hepatocyte-like cells obtained from stem cell differentiation and direct reprogramming have cells similar to mature liver cells morphology,function,and gene expression profile will have greater application prospects in the future.(3)3D suspension culture has excellent performance in terms of culture density and mass transfer efficiency,and has the advantages of easy amplification,convenient inoculation,and convenient access.It is the main development direction of in vitro 3D culture of hepatocytes.(4)Although the existing bioreactor system

关 键 词：生物人工肝 肝细胞 3D培养 增殖 规模化 生物反应器系统 实时在线 

分 类 号：R459.9[医药卫生—治疗学] R363[医药卫生—临床医学]