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[人膀胱癌原代培养及无血清培养法建立人膀胱癌细胞系]

[Primary culture of human bladder carcinomas and establishment of human bladder carcinoma cell line by serum-free culture].

作者信息

Ito H, Yamaguchi K, Kotake T, Matsuzaki O

出版信息

Nihon Hinyokika Gakkai Zasshi. 1989 Dec;80(12):1749-54. doi: 10.5980/jpnjurol1989.80.1749.

Abstract

It was possible to cultivate cells from bladder carcinoma tissues in 4 cases out of 6 without the overgrowth of fibroblast. A new human transitional cell carcinoma cell line (HAMT-1) was established in longterm tissue culture by using serum-free medium (BEM-841) which had been developed by us. The tissue for culture was taken from a 61-year-old Japanese male with grade 3 transitional cell carcinoma of the bladder. The microscopic features of the cell in cultures and of the tumors developed in nude mice resembled closely that of the original tumor. Electron microscopy of the cultured cells and the tumors developed in nude mice revealed characteristics of the epithelial origin of these cells with microvilli, junctional complex and scarce filament formation. Blood TPA level of the nude mouse with the transplanted tumor was equally high as that of the patient from whom the original tumor had been taken. The cells were anchorage independent in the serum-free medium but anchorage dependent in the medium containing 5% FCS. Anchorage dependency could not be restored by the addition of collagen and fibronectin. The doubling time of the cells were 18-20 hours. The chromosome counts of the cell line ranged from 59-78 with a modal count of 74.

摘要

6例膀胱癌组织中有4例成功培养出细胞,且无成纤维细胞过度生长。我们利用自行研发的无血清培养基(BEM - 841),通过长期组织培养建立了一种新的人膀胱移行细胞癌细胞系(HAMT - 1)。用于培养的组织取自一名61岁患膀胱3级移行细胞癌的日本男性。培养细胞及裸鼠体内形成肿瘤的显微镜特征与原发肿瘤极为相似。对培养细胞及裸鼠体内肿瘤进行电子显微镜检查,发现这些细胞具有上皮起源的特征,有微绒毛、连接复合体且细丝形成稀少。移植肿瘤的裸鼠血液中组织型纤溶酶原激活物(TPA)水平与原发肿瘤患者的水平一样高。细胞在无血清培养基中不依赖贴壁生长,但在含5%胎牛血清(FCS)的培养基中依赖贴壁生长。添加胶原蛋白和纤连蛋白无法恢复贴壁依赖性。细胞的倍增时间为18 - 20小时。该细胞系的染色体数在59 - 78之间,众数为74。

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