Pera M F, Blasco Lafita M J, Mills J
Int J Cancer. 1987 Sep 15;40(3):334-43. doi: 10.1002/ijc.2910400309.
The understanding of differentiation in human teratomas requires a better definition of the phenotype and developmental potential of the stem cells in these tumours. We describe the characterization of 6 new cell lines from human testicular teratomas which are representative of 3 distinct cell types. Cell lines GCT 27, GCT 35, and GCT 48, identified as embryonal carcinoma, comprise epithelial cells which express cytokeratin intermediate filaments and desmoplakins, as determined by indirect immunofluorescence microscopy. A minority of the cells also express vimentin. Most cells in these cultures show surface staining with monoclonal antibodies (MAbs) to stage-specific embryonic antigens SSEA 3 and SSEA 4 but not SSEA 1. Staining with MAb W6/32, which recognizes HLA A,B and C chains in the presence of beta-2 microglobulin, is not above background level. When injected into nude mice, GCT 27 cells form tumours consisting of embryonal carcinoma, somatic tissues, and cells positive in immunocytochemical assays for alphafoetoprotein (AFP) and human chorionic gonadotrophin (HCG); GCT 35 cells form embryonal carcinomas with foci of AFP and HCG-positive cells; and GCT 48 cells form embryonal carcinoma only. A second type of cell (GCT 72) displays some properties of rodent visceral endoderm. GCT 72 cells contain cytokeratin intermediate filaments, but not vimentin, and show very strong staining at cell borders with anti-desmoplakin I + II antibody. At the cell surface, GCT 72 cells express the epitopes recognized by antibodies to SSEA 3, SSEA 4, and SSEA 1; staining with W6/32 is negligible. Levels of AFP in supernatants from GCT 72 cultures are in excess of 500 KIU/I. The tumours formed following inoculation of nude mice with GCT 72 cells are solid yolk-sac tumours, with all cells strongly positive for AFP. A third cell type (GCT 44 and GCT 46), resembles in some ways rodent parietal endoderm. These cells uniformly coexpress keratin and vimentin intermediate filaments, but not desmoplakins. The determinants recognized by MAbs to SSEA 3, 4, or 1 are not detected on the majority of cells in these cultures. In striking contrast to the other teratoma lines, these cells can attach to untreated tissue culture plastic in serum-free medium and may be serially cultivated under these conditions. The tumours formed in nude mice by these 2 cell lines are yolk-sac carcinomas with endodermal sinus tumour histology. Thus, human testicular teratomas consist of epithelial cells which may be nullipotent, pluripotent or committed to extraembryonic endodermal differentiation.(ABSTRACT TRUNCATED AT 400 WORDS)
要了解人类畸胎瘤中的分化情况,需要更好地定义这些肿瘤中干细胞的表型和发育潜能。我们描述了从人类睾丸畸胎瘤中分离出的6种新细胞系的特征,它们代表了3种不同的细胞类型。细胞系GCT 27、GCT 35和GCT 48被鉴定为胚胎癌,由上皮细胞组成,通过间接免疫荧光显微镜检测,这些上皮细胞表达细胞角蛋白中间丝和桥粒斑蛋白。少数细胞也表达波形蛋白。这些培养物中的大多数细胞用针对阶段特异性胚胎抗原SSEA 3和SSEA 4的单克隆抗体(MAb)进行表面染色,但不表达SSEA 1。用识别β2微球蛋白存在时的HLA A、B和C链的MAb W6/32染色,未超过背景水平。当注入裸鼠体内时,GCT 27细胞形成的肿瘤由胚胎癌、体细胞组织以及在免疫细胞化学检测中对甲胎蛋白(AFP)和人绒毛膜促性腺激素(HCG)呈阳性反应的细胞组成;GCT 35细胞形成含有AFP和HCG阳性细胞灶的胚胎癌;GCT 48细胞仅形成胚胎癌。第二种细胞类型(GCT 72)表现出一些啮齿动物内脏内胚层的特性。GCT 72细胞含有细胞角蛋白中间丝,但不含有波形蛋白,并且用抗桥粒斑蛋白I + II抗体在细胞边界处显示出非常强的染色。在细胞表面,GCT 72细胞表达被针对SSEA 3、SSEA 4和SSEA 1的抗体识别的表位;用W6/32染色可忽略不计。GCT 72培养物上清液中的AFP水平超过500 KIU/I。用GCT 72细胞接种裸鼠后形成的肿瘤是实性卵黄囊瘤,所有细胞对AFP均呈强阳性。第三种细胞类型(GCT 44和GCT 46)在某些方面类似于啮齿动物的壁层内胚层。这些细胞均匀地共表达角蛋白和波形蛋白中间丝,但不表达桥粒斑蛋白。在这些培养物的大多数细胞上未检测到被针对SSEA 3、4或1的MAb识别的决定簇。与其他畸胎瘤细胞系形成鲜明对比的是,这些细胞可以在无血清培养基中附着于未处理的组织培养塑料上,并可以在这些条件下连续培养。这两种细胞系在裸鼠体内形成的肿瘤是具有内胚窦瘤组织学特征的卵黄囊癌。因此,人类睾丸畸胎瘤由上皮细胞组成,这些上皮细胞可能是全能性、多能性的,或者已定向于胚外内胚层分化。(摘要截短于400字)