摘要
细胞永生化是一项运用多种手段人为地将不同类型的外源性永生化基因导入目的细胞,打破细胞的正常分裂周期并使其具备无限增殖能力的技术,现已被广泛应用于不同动物原代细胞中。为了探究促使细胞永生化的几种常用方法如何介导细胞永生化,不同方法之间的相关因素有何关联和区别,以及这些方法如何应用到不同动物的细胞中,本文通过对几种促使细胞永生化的作用机制、方法及其在不同动物细胞(如猪单核细胞/巨噬细胞系、牛肠上皮细胞系、牦牛瘤胃上皮细胞系、鸡前脂肪细胞系、鸡胚胎成纤维细胞系、鸭肠上皮细胞系、兔黑色素细胞系和斑节虾淋巴细胞系等)中的应用现状进行概述,发现细胞永生化主要涉及端粒与端粒酶激活、病毒基因[如人类疱疹病毒(EBV)、猴空泡病毒40(SV40)和人乳头瘤病毒(HPV)基因]的转染整合,以及运用Cre-LoxP介导的重组系统对细胞进行可回复性永生化等多个方法,且其核心都是对细胞端粒酶活性的调控。此外,不同动物细胞可通过上述多种方法实现永生,但是也有极少部分细胞受其自身性质的影响,只可选择某一特定方法。因此,对不同动物原代细胞永生化方法及其应用进行研究,不仅可为建立具备功能性的永生化细胞系提供理论依据,也为其在基础分子生物学研究和生物医药领域更广泛的潜在应用提供科学资料。
Cell immortalization is a technique that artificially introduces various exogenous immortalizing genes into target cells,disrupts the normal cell division cycle and endows them with unlimited proliferation potential.It has been widely applied in primary cells from different animal species.To explore how several commonly used methods induce cell immortalization,the correlation and differences among these methods,as well as their application to cells from different animal species,this article provides an overview of the mechanisms and methods for inducing cell immortalization,and their current applications in different animal cells,such as porcine alveolar macrophage line,bovine intestinal epithelial cell line,yak rumen epithelial cell line,chicken preadipocyte line,chicken embryo fibroblast line,duck intestinal epithelial cell line,rabbit melanocyte line,and lymphocyte line from shrimp.This article revealed that cell immortalization mainly involves the regulation of telomeres and telomerase activation,transfection and integration of viral gene,such as human herpesvirus(EBV),simian virus 40(SV40),and human papillomavirus(HPV)genes,as well as the use of Cre-LoxP-mediated recombination system for reversible cell immortalization,with the core being the regulation of cellular telomerase activity.Additionally,different animal cells can achieve immortalization through the aforementioned methods,but some cells may be influenced by their intrinsic characteristics,resulting in the selection of specific methods.Therefore,studying the methods and applications of immortalization in primary cells from different animals not only provides a theoretical basis for establishing functional immortalized cell lines but also offers scientific data for broader potential applications in basic molecular biology research and the biopharmaceutical field.
作者
侯碧巍
董书餐
刘德武
孙宝丽
李耀坤
郭勇庆
邓铭
柳广斌
HOU Bi-wei;DONG Shu-can;LIU De-wu;SUN Bao-li;LI Yao-kun;GUO Yong-qing;DENG Ming;LIU Guang-bin(College of Animal Science,South China Agricultural University,Guangzhou 510642,China)
出处
《中国兽医杂志》
CAS
北大核心
2023年第8期97-103,共7页
Chinese Journal of Veterinary Medicine
基金
广东省自然科学基金项目(2021A1515010636)
广东省现代农业产业技术体系项目(2022KJ127)
韶关市仁化县牛羊产业园山羊养殖技术服务项目(h20220665)。
关键词
原代细胞
细胞永生化
端粒酶
病毒基因
可回复性永生化
primary cells
cell immortalization
telomerase
viral genes
reversible immortalization
