大肠癌细胞的原代培养方法  

The Study on the Primary Culture Methods of Human Colorectal Cancer Cells

作　　者：杨磊磊[1] 戴岳楚[1] 董米连[1] 朱敏[1] 林雪飞[1] 廖伟[2] 梅统[2] 

机构地区：[1]浙江省临海市温州医学院附属浙江省台州医院,临海317000 [2]浙江省温州市温州医学院,温州325000

出　　处：《中国中西医结合外科杂志》2012年第4期341-345,共5页Chinese Journal of Surgery of Integrated Traditional and Western Medicine

基　　金：浙江省医学会临床科研基金项目(2009ZYC50);台州市科技计划项目(2010-26-48)

摘　　要：目的:探讨大肠癌细胞体外分离培养方法,建立合理、高效、稳定的短期大肠癌细胞原代培养模式。方法:取60例大肠癌(T3或T4,且CT示肿瘤直径>2.0 cm)的新鲜肿瘤组织,随机分为黏膜层取材组与浆膜层取材租,分别采用组织块法、机械分离法、胰酶消化法、胶原酶消化法进行体外培养大肠癌细胞,对不同方法及培养条件进行比较并加以改进。结果:黏膜层取材组优于浆膜层取材组(P<0.05),胶原酶消化法优于胰酶消化法(P<0.05),胰酶消化法优于机械分离法(P<0.05),胶原酶消化法是最优的人类大肠癌细胞原代培养方法,细胞培养成功率达到66.7%。结论:通过黏膜层取材的胶原酶消化法可成功建立较为合理、高效、稳定的短期大肠癌细胞原代培养模式。Objective To investigate the human colorectal cancer cells how to be cultured in vitro methods, then establish a rational, efficient and stable short-term primary culture model of colorectal cancer cells. Methods 60 cases of colorectal cancer （T3 or T4, and CT showed： tumor size〉2.0 cm） of fresh tumor tissue which were diagnosed in the hospital were randomly divided into 2 groups,namely the mucous layer and serosal layer derived group. Tissue were divided in these vitro methods to cultivate human colorectal cancer cells ,then culture conditions on the different methods were compared and improved.These methods include tis sue nubbles cultivation, mechanical separation, trypsin digestion, collagenase digestion and cultured. Results The success rate of the mucous layer derived group was better than the mucosal serosal layer derived group （P 〈 0.05）. Col- lagenase digestion method was superior to trypsin digestion method （P 〈 0.05）. Trypsin digestion method was bet- ter than mechanical separation method to achieve the target （P 〈 0.05）.Among these methods, collagenase diges- tion method in which human colorectal cancer cells is the best method for primary culture.The success rate of cell cultivation could be 66.7%. Conclusion The more rational, efficient and stable short-term primary cul- ture model of colorectal cancer cells has been established,which is to draw through the mucous layer,in opti- mized collagenase digestion method.

关 键 词：大肠癌细胞 原代培养 鉴定 黏膜层 胶原酶消化法 

分 类 号：R735.34[医药卫生—肿瘤] Q813.11[医药卫生—临床医学]