中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (2): 273-276.doi: 10.3969/j.issn.1673-8225.2012.02.019

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

改良Alamar Blue法检测结直肠微小组织块的活力***☆

甘嘉亮1，高  枫1，曹云飞1，廖  存2，黄家豪2，唐双意3   

  1. 广西医科大学第一附属医院，1结直肠肛门外科，  3药剂科，广西壮族自治区南宁市530021；2广西医科大学研究生学院，广西壮族自治区南宁市 
    530021
  • 收稿日期:2011-09-01 修回日期:2011-12-02 出版日期:2012-01-08 发布日期:2012-01-08
  • 通讯作者: 高枫，博士，教授，广西医科大学第一附属医院结直肠肛门外科，广西壮族自治区南宁市 530021 gaofeng0771@126.com
  • 作者简介:甘嘉亮☆，男，1972年生，广西壮族自治区贵港市人，汉族，广西医科大学在读博士。 gjl259@126.com
  • 基金资助:

    广西教育厅科研资助项目，课题名称：CEA阳性和CEA阴性结直肠癌蛋白组学比较研究”(201012MS039)和“蛋白组学技术及其在结直肠癌中的应用研究”(201012MS041)；广西2011年研究生教育创新计划项目(2011105981002D34)。

Viability of tissue fragments in colon and rectum detected using modified Alamar Blue assay  

Gan Jia-liang1, Gao Feng1, Cao Yun-fei1, Liao Cun2, Huang Jia-hao2, Tang Shuang-yi3   

  1. 1Department of Colorectal and Anal Surgery,  3Department of Pharmacy, First Affiliated Hospital of Guangxi Medical University, Nanning  530021, Guangxi Zhuang Autonomous Region, China;
    2Postgraduate College of Guangxi Medical University, Nanning  530021, Guangxi Zhuang Autonomous Region, China
  • Received:2011-09-01 Revised:2011-12-02 Online:2012-01-08 Published:2012-01-08
  • Contact: Gao Feng, Doctor, Professor, Department of Colorectal and Anal Surgery, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China gaofeng0771@126.com
  • About author:Gan Jia-liang☆, Studying for doctorate, Department of Colorectal and Anal Surgery, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China gjl259@126.com
  • Supported by:

    the Scientific Research Foundation of Education Bureau of Guangxi Zhuang Autonomous Region, No. 201012MS039*, 201012MS041*; Innovation Project of Guangxi Graduate Education No.2011105981002D34*

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摘要：

背景：Alamar Blue法是测定组织细胞活性的方法之一，目前多数应用在细胞系活力测定方面，尚未见应用于微小组织块活力的评价。
目的：采用改良Alamar Blue法评价结直肠正常黏膜组织块和癌组织块的活力。
方法：以细胞活力测定的Alamar Blue法为基础，应用单位质量吸光度来评价结直肠正常黏膜组织块和癌组织块的活力。根据检测试剂盒说明书和预实验结果，实验选择0，6，12，18，24，30，36 h作为检测时间点，每一例标本分为7组，设空白对照孔。
结果与结论：单位质量吸光度能够随培养时间的推移，很好地反映组织块增殖活力的变化趋势，结直肠正常黏膜组织块和癌组织块培养12~24 h达到良好的增殖状态，证实改良Alamar Blue法是评价结直肠微小组织块灵敏有效的新型检测方法之一。

关键词: 微小组织块, Alamar Blue法, 增殖, 活力, 组织构建

Abstract:

BACKGROUND: Alamar Blue assay is one of the common methods to evaluate histic or cellular viability. It is widely used to detect cell viability at present. However, using Alamar Blue assay to detect the viability of tissue fragments has not been reported.
OBJECTIVE: To detect the viability of the regular tissue fragments and cancer tissue fragments in the mucous membrane of colon and rectum using modified Alamar Blue assay.
METHODS: Based on cell viability detect assay of Alamar Blue, the viability of the regular tissue fragments and cancer tissue fragments in the mucous membrane of colon and rectum was assessed using absorbance per unit mass. The detecting time was chosen at 0, 6, 12, 18, 24 , 30 and 36 hours according to the specification of the Alamar Blue assay and the pre-experiment results. Every specimen was divided into seven groups and the blank control hole was established.
RESULTS AND CONCLUSIONS: Absorbance per unit mass reflected the change tendency of proliferation activity in tissue fragments as the culture time went on. Both regular tissue fragments and cancer tissue fragments in the mucous membrane of colon and rectum reached the best proliferative state at 12-24 hours after culture. It confirms that the modified Alamar Blue assay is a sensitive and effective way to evaluate the viability of colorectal tissue fragments.

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