黑色素瘤细胞系HME1的建立及其生物学特性分析

Establishment and characterization of a melanoma cell line HME1

目的 :建立一株黑色素瘤细胞系 ,并分析其体外生物学特性。方法 :取黑色素瘤活检组织进行组织块法原代培养 ,待长满 90 %后 ,消化传代培养并进行生长动力学、形态学及致瘤性等生物学特性鉴定。结果 :该细胞系已在体外培养生存 1 8个多月 ,传 90余代。其生物学特性如下 :该细胞系接种至裸鼠后可致瘤 ,其移植瘤细胞形态与原实体瘤相似 ;检测第 2 0代细胞生长曲线 ,其倍增时间为 5 6 .9h ;第 2 0代细胞软琼脂集落形成率平均为 1 9.1 % ;染色体核型分析为非整倍体 ,众数为 92条 ;透射电镜下可观察到细胞表面有丰富的微绒毛、核糖体及黑色素颗粒 ;SP免疫组化分析显示 ,细胞有HMB 4 5表达。结论 :该黑色素瘤细胞经体外培养后已永生化 ,形成了连续传代细胞系 。

Objective To establish a melanoma cell line and identify its characteristics in vitro. Methods The tissues from biopsies of melanoma were performed primary culture. After growing to 90% confluence, the cells were detached and transferred to another flask for subculture, and then we identified their characteristics including growth kinetic, morphology and tumorigenecity. Results This cell line was cultivated for more than 90 times. Its characteristics were as follows:The pathological morphology of the tumor transplanted in nude mice were similar to that of the original lesion;The growth curves of the 20th passage were determined,and the population doubling time calculated was 56.9 h; The cloning efficiency in soft agar was 19.1%;Karyo type analysis showed aneuploidy with the modal chromosomal number 85 102;Electronmicroscopical observation showed that there were rich microvilli on the surface of the cells, abundant ribosomes and melanoid grain;SP immunohistochemical staining showed that the cells expressed HMB 45. Conclusion The Melanoma cells were immortalized after being cultured in vitro and a new melanoma cell line was established.