8株人黑色素瘤细胞系的建立及其生物学特性研究

目的：从人黑色素瘤组织建立一系列黑色素瘤细胞系，命名为黑色素瘤细胞系1-8号（HME1-8），并研究鉴定其体外细胞生物学与免疫学特性。方法：取黑色素瘤组织进行组织块培养法，待长满90%汇合面后消化传代培养，并进行形态学、生长动力学、免疫学及致瘤性等生物学特性研究。结果：该系列黑色素瘤细胞系已在体外培养生存1-2年，传60-100余代，其生物学特性显示：①这些细胞系体外生长的倍增时间为34.2-59.5h；②软琼脂集落形成率平均值在8.6%-26.2%之间；③接种至裸鼠后具有较高的致瘤性；④染色体核型分析为非整倍体，众数为64-117条；⑤ 透射电镜下可观察到细胞含丰富的核糖体及黑色素颗粒；⑥免疫组化分析显示细胞浆内含大量阳性棕色黑色素颗粒；⑦肿瘤抗原检测显示8株黑色素瘤细胞系的Melan-A抗原阳性率为75%, MAGE-1阳性率为50%, MAGE-2阳性率为37.5%，MAGE-3阳性率为62.5%。结论：8株黑色素瘤细胞系经体外长期培养后已永生化，形成了连续传代细胞系，具有明显的黑色素瘤细胞恶性表型特征。

Characteristics of eight established human melanoma cell lines

AIM: Eight melanoma cell lines were established from human melanoma specimen and designated as HME 1-HME 8, and their characteristics of cell biology and immunology in vitro were studied. METHODS: The tissues from biopsies of human melanoma were cultured in RPMI 1640 media. After growing to 90% confluence, the cells were detached for subculture and then their characteristics, including morphology, growth kinetic, tumor antigen and tumorigencity in nude mice were studied. RESULTS: These cell lines have been passaged more than 100 times within 2 years. Their characteristics demonstrated: ① The population doubling time calculated in the log phase of growth were 34.2-59.5 h. ② The cloning efficiency in soft agar averaged 8.6%-26.2%. ③ The melanoma cells transplanted in nude mice were high tumorigencity. ④ Karyo-type analysis showed that aneuploidy with the modal chromosomal number 64-117. ⑤ Rich ribosomes and melanoid grains in the cytoplasm were observed under electronic microscope. ⑥ Immunohistochemistry analysis showed that there were a lot of brown grains in the cytoplasm. ⑦ Detection of tumor-antigen demonstrated that melan-A antigen was 75.0%, MAGE-1 was 50.0%, MAGE-2 was 37.5%, MAGE-3 was 62.5%, respectively, for eight melanoma cell lines. CONCLUSION: The melanoma cells have immortalized after being cultured in vitro and has specific characteristics of malignant melanoma.