Pietsch T, Scharmann T, Fonatsch C, Schmidt D, Ockler R, Freihoff D, Albrecht S, Wiestler O D, Zeltzer P, Riehm H
Department of Pediatric Hematology and Oncology, Medical School Hannover, Germany.
Cancer Res. 1994 Jun 15;54(12):3278-87.
Medulloblastoma (MB) represents the most frequent malignant brain tumor of childhood but only a few cell lines and animal models of this primitive neuroectodermal tumor (PNET) have thus far been established. Using specific cell culture conditions, we were able to derive four human MB cell lines (MHH-MED-1-4) as well as a cell line from a spinal PNET (MHH-PNET-5). The four MB cell lines grew in suspension as floating cell aggregates or as slightly adherent cells. They consisted of undifferentiated cells that did not express markers of late neuronal or glial lineages such as neurofilaments or glial fibrillary acidic protein. They also lacked expression of major histocompatibility complex class I or II antigens on the cell surface. All four MB lines were positive for vimentin and neuron-specific enolase, whereas synaptophysin, neural cell adhesion molecule, galactocerebroside, GD2, GD3, and the A2B5 antigen were expressed inconsistently. In contrast, MHH-PNET-5 grew as adherent monolayer and expressed major histocompatibility complex class I antigen. By cytogenetic analysis, the lines were near diploid with clonal aberrations. The MB lines showed no losses of chromosome arm 17p by either cytogenetic or microsatellite analyses. The cell line MHH-MED-2 exhibited double minute chromosomes, amplification of the c-myc gene, and overexpression of c-myc mRNA and protein. N-myc, p53, and Rb protein expression were unaltered. All four continuously passaged MB cell lines and the MHH-PNET-5 line were xenotransplanted s.c. into athymic mice; three of four MB lines and the spinal PNET line gave rise to tumors. These cell lines will be useful tools for biological and preclinical studies on PNETs.
髓母细胞瘤（MB）是儿童期最常见的恶性脑肿瘤，但迄今为止，这种原始神经外胚层肿瘤（PNET）仅建立了少数细胞系和动物模型。通过特定的细胞培养条件，我们成功获得了4个人类MB细胞系（MHH-MED-1-4）以及1个来自脊髓PNET的细胞系（MHH-PNET-5）。这4个MB细胞系以悬浮状态生长，形成漂浮的细胞聚集体或轻度贴壁细胞。它们由未分化细胞组成，不表达晚期神经元或神经胶质谱系的标志物，如神经丝或胶质纤维酸性蛋白。它们在细胞表面也缺乏主要组织相容性复合体I类或II类抗原的表达。所有4个MB细胞系波形蛋白和神经元特异性烯醇化酶呈阳性，而突触素、神经细胞黏附分子、半乳糖脑苷脂、GD2、GD3和A2B5抗原的表达则不一致。相比之下，MHH-PNET-5以贴壁单层形式生长，并表达主要组织相容性复合体I类抗原。通过细胞遗传学分析，这些细胞系接近二倍体，伴有克隆性畸变。无论是细胞遗传学分析还是微卫星分析，MB细胞系均未显示17号染色体短臂缺失。细胞系MHH-MED-2表现出双微体染色体、c-myc基因扩增以及c-myc mRNA和蛋白的过表达。N-myc、p53和Rb蛋白表达未改变。所有4个连续传代的MB细胞系和MHH-PNET-5细胞系均皮下移植到无胸腺小鼠体内；4个MB细胞系中的3个以及脊髓PNET细胞系均形成了肿瘤。这些细胞系将成为PNET生物学和临床前研究的有用工具。
