中国组织工程研究 ›› 2017, Vol. 21 ›› Issue (9): 1324-1328.doi: 10.3969/j.issn.2095-4344.2017.09.003

• 肿瘤干细胞 cancer stem cells • 上一篇    下一篇

基于“单细胞移植-成瘤性验证”模式鉴定肝癌干细胞

贾  微,岑妍慧,包  鹃,杨  瑞,何国珍,吴晓君,李熠毅   

  1. 广西中医药大学,广西壮族自治区南宁市  530222
  • 出版日期:2017-03-28 发布日期:2017-03-31
  • 通讯作者: 岑妍慧,博士,副教授,广西中医药大学,广西壮族自治区南宁市 530222 并列通讯作者:杨瑞,硕士,讲师,广西中医药大学,广西壮族自治区南宁市 530222
  • 作者简介:贾微,男,1982年生,辽宁省沈阳市人,汉族,硕士,讲师,研究方向为中医药、干细胞的体外分离及培养等方面。 并列第一作者:包鹃,广西中医药大学,广西壮族自治区南宁市 530222
  • 基金资助:

    国家自然科学基金项目(81503406);广西高校科研课题(ZD2014068);广西高校青年教师能力提升计划(KY2016YB218,KY2016YB224);广西壮族自治区卫生厅中医药科技专项课题(GZPT13-04,GZBZ16-07,GZLC16-23),广西中医药大学学生科研训练课题(2014DXS02)

A preliminary establishment of the “single-cell-transplant-umorigenic-test”model for identifying liver cancer stem cells

Jia Wei, Cen Yan-hui, Bao Juan, Yang Rui, He Guo-zhen, Wu Xiao-jun, Li Yi-yi   

  1. Guangxi University of Chinese Medicine, Nanning 530222, Guangxi Zhuang Autonomous Region, China
  • Online:2017-03-28 Published:2017-03-31
  • Contact: Cen Yan-hui, M.D., Associate professor, Guangxi University of Chinese Medicine, Nanning 530222, Guangxi Zhuang Autonomous Region, China Corresponding author: Yang Rui, Master, Lecturer, Guangxi University of Chinese Medicine, Nanning 530222, Guangxi Zhuang Autonomous Region, China
  • About author:Jia Wei, Master, Lecturer, Guangxi University of Chinese Medicine, Nanning 530222, Guangxi Zhuang Autonomous Region, China Bao Juan, Guangxi University of Chinese Medicine, Nanning 530222, Guangxi Zhuang Autonomous Region, China Jia Wei and Bao Juan contributed equally to this work.
  • Supported by:

    the National Natural Science Foundation of China, No. 81503406; University Research Projects in Guangxi, No. ZD2014068; Project of Improving the Basic Ability of Young Teachers in Universities in Guangxi, No. KY2016YB218, KY2016YB224; Special Scientific Project of Traditional Chinese Medicine supported by Health Department of Guangxi Zhuang Autonomous Region, No. GZPT13-04, GZBZ16-07, GZLC16-23; and Student Research Training Project of Guangxi Medical University, No. 2014DXS02

摘要:

 

文题释义:

单细胞移植技术:经过实验摸索,在显微操作法的基础上改进,利用最小号的静脉注射静脉注射针及微量移液器制作了单细胞分离种植装置,该技术分离单细胞具有取材容易、制作简单、操作直观可靠、分离成功率高的特点。而最后经过这一技术的分离,单细胞/孔的成功率达到了96.4%。

 

 
摘要
背景:目前尚无较有效的肝癌干细胞的鉴定方法。
目的:探索更为简单有效并适用于鉴定肝癌肿瘤细胞株中干细胞的方法。
方法:首先利用单细胞分离技术获得单个细胞,逐个种入96孔板中,经筛选获得单细胞克隆(即成瘤性克隆)来源的细胞亚株,细胞亚株移植到裸鼠前肢腋下观察其成瘤作用。 

结果与结论:利用单细胞分离技术获得单细胞/孔的孔数为371个,成功率为96.4%;按照单细胞克隆的生长情况,筛选获得的来源于成瘤性克隆的细胞亚株移植到裸鼠前肢腋下,成瘤率为100%。结果证实“单细胞移植-成瘤性验证”具有鉴定肝癌干细胞的有效性,为后续的研究奠定技术和方法基础。

 

中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程
ORCID:
0000-0002-6020-0616(Cen Yan-hui)

关键词: 干细胞, 肿瘤干细胞, 单细胞移植-成瘤性, 肝癌干细胞, 干细胞鉴定, HepG2肝癌细胞, 裸鼠, 细胞移植, 国家自然科学基金

Abstract:

BACKGROUND: There is no effective method to identify liver cancer stem cells until now, which has become one of the major challenges in this field.
OBJECTIVE: To explore a more effective and suitable way for the identification of liver cancer stem cells in hepatocellular carcinoma cell lines.
METHODS: Firstly, the single cell separation technique was used to obtain single cells, which were seeded into 96-well plates one by one. Secondly, cell sublines derived from single cell colonies (tumorigenic colonies) were selected and obtained. At last, the cells from these clones were transplanted into the forelimb armpits of nude mice to observe the tumorigenic ability.
RESULTS AND CONCLUSION: The number of holes of single cells obtained was 371 by the single cell separation technique, and the success rate was 96.4%. According to the growth of cell clone, tumorigenic colonies were selected to be transplanted to the forelimb armpits of nude mice, and the tumor formation rate of the colonies was 100%. The identification model of “single-cell-transplant-tumorigenic-test” for liver cancer stem cells is confirmed to be preliminarily established, which lays the technology and methodological basis for the follow-up research.

 

中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程

Key words: Hep G2 Cells, Neoplastic Stem Cells, Tissue Engineering

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