人葡萄膜黑色素瘤的细胞培养经验及鉴定

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-100，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-100，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。

Culture of human uveal melanoma

Objective: To study the culture method of human uveal melanoma in vitro and the characteristics of uveal melanoma cells. Methods: 3 ways were used to prepare cells, the mechanical, chemical, and tissue culture method. Cell morphology was observed with inverse microscope or HE stain. The expressions of melanoma associated antigen HMB45, S-100, and NSE were checked with immunocytochemical stain, ultra structure of cell was checked with electron microscope. Results: Mechanical separation could get enough living melanoma cells than the other two ways. Cell types were epithelia and spindle; HMIMS, S-100 and NSE were positive. A prominent nucleus, diffuse nuclear chromatin, different stages and irregular morphology of melanosome development were observed by electron microscope. Conclusions: Mechanical separation seems the best way for uveal melanoma cell culture in vitro. With immunocytochemical staining and electron microscopy the resources and purity of cells can be identified.