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目的 :探讨人葡萄膜黑色素瘤细胞体外培养的方法及生物学鉴定的研究。方法 :分别用机械取材法、化学取材法及组织块培养法 ,体外培养人葡萄膜黑色素瘤细胞。通过倒置显微镜下观察细胞形态及生长特性 ,收集第三代融合的细胞经原位固定后作 HE染色、 HMB45 ,S- 10 0 ,神经元特异性烯醇化酶 (NSE)免疫组化染色 ,电子显微镜鉴定培养细胞。结果 :三种取材方法中 ,机械分离的肿瘤细胞活性好、繁殖快 ,细胞呈梭型或类上皮型 ,胞核透明呈梭形或椭圆形 ,胞浆富含黑色素颗粒。HMB45 ,S- 10 0 ,NSE呈阳性反应。透射电镜发现肿瘤细胞内不同发育阶段的黑色素小体及大小不等的黑色素颗粒。结论 :机械取材法是获取人葡萄膜黑色素瘤细胞体外培养的最佳方法。用免疫组化及电镜可以鉴定细胞来源及纯度
Objective: To study the method and biological identification of human uveal melanoma cells cultured in vitro. Methods: Human uveal melanoma cells were cultured in vitro by using the method of mechanical drawing, chemical drawing and tissue culture respectively. The morphology and growth characteristics of the cells were observed under an inverted microscope. The cells in the third generation were collected and fixed for HE staining, HMB45, S-10 0, NSE immunohistochemistry, Microscopic identification of cultured cells. Results: Among the three methods, the tumor cells isolated mechanically had good activity and proliferated quickly. The cells were shuttle type or epithelioid type. The nuclei were fusiform or oval in shape, and the cytoplasm was rich in melanin granules. HMB45, S-10 0, NSE positive reaction. Transmission electron microscopy revealed different stages of development of melanosomes in tumor cells and melanin particles of varying sizes. Conclusion: The mechanical method is the best way to obtain human uveal melanoma cells cultured in vitro. Immunohistochemistry and electron microscopy can identify the cell source and purity