干细胞扩增及肝向分化过程中相关指标及其检测方法  

Stem cell expansion and hepatic differentiation:measurement of related indicators and detection methods

作　　者：杨嘉屹 张广浩[1,2] 张丞 李柯[1,2] 霍小林 吴昌哲[1,2,3] Yang Jiayi;Zhang Guanghao;Zhang Cheng;Li Ke;Huo Xiaolin;Wu Changzhe(Beijing Key Laboratory of Bioelectromagnetics,Institute of Electrical Engineering,Chinese Academy of Sciences,Beijing 100190,China;University of Chinese Academy of Sciences,Beijing 100049,China;Zhujiang Hospital,Southern Medical University,Guangzhou 510282,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510282

出　　处：《中国组织工程研究》2020年第25期4039-4045,共7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;广东省自然科学基金研究团队项目(2014A030312013),项目参与人:吴昌哲。

摘　　要：背景:干细胞在组织工程学、再生医学、细胞治疗以及药物研究方面具有巨大的应用前景。近年来,以生物人工肝为例的应用研究需要大规模扩增出高质量的干细胞并最终定向分化为肝(样)细胞。如何通过此途径高效获得一致性、功能性完好并且标志物均匀表达的分化细胞,是需要解决的关键问题。目的:对干细胞扩增及肝向分化过程相关检测指标与检测方法进行综述,总结已应用于在线检测的指标与方法,讨论部分指标与方法应用于在线检测的局限性,并对未来有望应用于在线检测的指标和检测技术进行展望。方法:由第一作者检索1990至2019年PubMed数据库、Web of Science核心合集、Elsevier数据库、中国知网等收录的与干细胞扩增及肝向分化过程相关的文献。英文检索词为“stem cells,expansion,hepatic differentiation,monitoring,real-time online”,中文检索词为“干细胞,扩增,肝向分化,监测,实时在线”,共计检索93篇文章,最终筛选了符合标准的53篇进行综述。结果与结论:干细胞扩增及肝向分化过程的相关检测指标与检测方法众多,其中扩增过程的检测指标主要涉及干细胞多能性的保持、代谢活性、存活率以及转癌趋势。肝向分化过程因细胞种类而异,其中多能干细胞和中胚层谱系成体多能干细胞要先向内胚层定向分化,再分化为成熟肝(样)细胞,而以肝干/祖细胞为主的内胚层谱系成体多能干细胞可直接分化为成熟肝(样)细胞。分化过程不同阶段细胞的特异性标志物、存活率以及代谢活性是检测的重点。目前,基于大型生化检测分析设备的检测方法虽然可靠性高,但面临实时性差、成本高、难以集成小型化等问题。未来,干细胞扩增及肝向分化过程中关键检测指标的筛选、检测标准的量化以及自动化在线检测的实现,需要重点研究。BACKGROUND:Stem cells have wide application prospects in tissue engineering,regenerative medicine,cell therapy and drug research.In recent years,the applied research,such as bioartificial liver,requires large-scale and high-quality stem cells by expansion,and final obtains hepatocytes(hepatocyte-like cells)by directional differentiation.Through this way,how to efficiently obtain the differentiated cells,with excellent consistency,powerful function and uniform expression of markers,are the key problems to be solved.OBJECTIVE:To review the detection indicators and detection methods related to the process of stem cell expansion and hepatic differentiation,summarize the indicators and methods that have been applied to online detection,discuss the limitations of some indicators and methods applied to online detection,and prospect the indicators and detection technologies expected to be applied to online detection in the future.METHODS:PubMed,Web of Science,Elsevier and CNKI databases were retrieved for the articles concerning stem cell expansion and hepatic differentiation published from 1990 to 2019.The literature related to stem cell expansion and hepatic differentiation was collected.The keywords were“stem cells;expansion;hepatic differentiation;monitoring;real-time online”in English and Chinese,respectively.A total of 93 articles were searched,and finally 53 eligible articles were selected and reviewed.RESULTS AND CONCLUSION:There are many detection indicators and detection methods for stem cell expansion and hepatic differentiation.Among them,the detection indicators of the expansion process are mainly related to the maintenance of pluripotency,metabolic activity,survival rate,and cancer transformation trend of stem cells.The process of hepatic differentiation varies with cell types,in which,the pluripotent stem cells and the mesodermal lineage adult pluripotent stem cells should differentiate into the endoderm-oriented cells and then the mature hepatocytes(hepatocyte-like cells),while the endodermal lineage

关 键 词：生物人工肝 干细胞 肝(样)细胞 细胞扩增 细胞分化 实时在线 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学]