人大肠癌细胞体外分离与培养  

Separation and Culture of Human Colorectal Cancer Cells in Vitro

作　　者：张毅强[1] 程牛亮[2] 

机构地区：[1]长治医学院生物化学教研室,046000 [2]山西医科大学

出　　处：《长治医学院学报》2009年第3期168-171,共4页Journal of Changzhi Medical College

摘　　要：目的:探讨人大肠癌细胞体外分离培养的方法。方法:采用组织块法和Ⅳ胶原酶消化法体外培养人大肠癌细胞,HE染色、免疫细胞化学判断细胞的组织来源、TEM观察细胞形态,MTT法绘制细胞生长曲线。结果:采用组织块法细胞培养成功,HE染色显示核大蓝染,细胞角蛋白鉴定胞浆黄染,细胞角蛋白阳性;透射电镜下,见细胞内丰富的线粒体、表面微绒毛、细胞间连接复合体结构;细胞生长曲线为"S"。结论:组织块法原代培养细胞成功率高,需注意在取材、分离操作过程中保持无菌,所培养的细胞进行初步鉴定证明为大肠癌细胞。Objective ： To investigate methods of separation and culture of human colorectal cancer cells in vitro. Methods ： Human colorectal cancer cells are cultured by explant method and Ⅳ collagenase digestion; HE stain, Immunocytochemistry identify tissue original, and transmission electron microscope TEM were used to view cells morphous; MTT assay was usedto draw cell growth curve. Results： HE staining showed karyomegaly and blue - stained colorectal cancer cells. Immunocytochemistry staining showed （cytoderatin）CK was positive, cytoplasm was yellow; TEM showed big cell nuclear, abundance in mitochondria, cell surface mi- crovillus and cell - cell junctional complex; Cell growth curve showed sigmoid curve. Conclusion： Explant method has a high achievement ratio. Explant should be kept asepsis during drawing materials and isolation. Cultured cells shluld be identified as colorectal cancer cells initially.

关 键 词：大肠癌 体外培养 原代培养 

分 类 号：R373[医药卫生—病原生物学]