Mohammadpour H, Hall M R, Pardini R S, Khaiboullina S F, Manalo P, McGregor B
Department of Veterans Affairs Medical Center, Reno, Nevada, 89520, USA.
J Surg Res. 1999 Apr;82(2):146-50. doi: 10.1006/jsre.1998.5544.
Techniques for creation of colon carcinoma epithelial cells lines in long-term culture have been available for years, but these techniques have involved mechanical or enzymatic methods to separate epithelial cells from surrounding tissues. While this practice has been intermittently successful, the effect of these traumatic methods on long-term cellular behavior is unknown. Samples of colon carcinoma from patient volunteers were subjected to serial nonenzymatic disruptions of carcinoma cells from surrounding fibrous tissues. Cells were collected, allowed to proliferate, and then tested for their epithelial characteristics (mucin, vimentin, cytokeratin, colon-specific antigen, carcinoembryonic antigen) by immunohistochemistry and flow cytometry. Growth characteristics were determined by phase-contrast microscopy, multiple passage, and freeze/thaw effects. Tumorigenicity was proven in nude mice. Of 11 initial attempts, three resulted in stable long-term culture lines of cells which are demonstrated to behave similarly to the original tumors from which they were derived. This technique adds another reliable in vitro tool for the study of colon carcinoma.
多年来一直有在长期培养中创建结肠癌细胞系的技术，但这些技术涉及机械或酶法将上皮细胞与周围组织分离。虽然这种做法偶尔会成功，但这些创伤性方法对细胞长期行为的影响尚不清楚。对来自患者志愿者的结肠癌样本进行了将癌细胞与周围纤维组织进行连续非酶破坏的操作。收集细胞，使其增殖，然后通过免疫组织化学和流式细胞术检测其上皮特征（粘蛋白、波形蛋白、细胞角蛋白、结肠特异性抗原、癌胚抗原）。通过相差显微镜、多次传代和冻融效应确定生长特性。在裸鼠中证实了致瘤性。在11次初始尝试中，有3次产生了稳定的长期细胞培养系，这些细胞系表现出与它们所源自的原始肿瘤相似的行为。这项技术为结肠癌研究增加了另一种可靠的体外工具。
