Vinores S A, Herman M M, Katsetos C D, May E E, Frankfurter A
Wilmer Ophthalmological Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287-9289.
Histochem J. 1994 Aug;26(8):678-85. doi: 10.1007/BF00158293.
The D283 Med human medulloblastoma cell line and primary explants of five surgically excised medulloblastomas were cultured using a three-dimensional Gelfoam matrix system. The cultures were evaluated immunohistochemically for a series of antigenic determinants associated with neuronal or glial differentiation. Focal immunolocalization of class III beta-tubulin, microtubule-associated protein 2 (MAP2), and to a lesser degree tau, was demonstrated in all cultures. Class III beta-tubulin isotype, MAP2, and tau protein were also detected by immunoblot in Gelfoam matrix cultures, monolayer cultures, and suspension cultures of D283 Med cells. Staining for neurofilament protein epitopes was highly variable, even among different cultures derived from the same original tumour, but time-dependent changes in neurofilament protein, which may have reflected neuronal differentiation, were not consistently shown. Widespread gamma-enolase and focal synaptophysin reactivities were visualized in all cultures, but no S-antigen staining was detected. Leu 7 labelling was variably present in half of the cultures of D283 Med cells, but was more abundant in explants derived from four of the five original tumours. Vimentin was consistently found in D283 Med cultures at all time points. No immunoreactivity for glial fibrillary acidic protein was detected in the D283 Med cell line. Conversely, staining for this protein was demonstrated in scattered astrocytic cells in the surgical specimens of all five medulloblastomas. Concomitant with increased time in culture, three of the primary tumours displayed increased numbers of glial fibrillary acidic protein-positive cells when cultured in the Gelfoam system, but the other two tumours had a minimal astrocytic component.(ABSTRACT TRUNCATED AT 250 WORDS)
使用三维明胶海绵基质系统培养D283 Med人髓母细胞瘤细胞系以及5例手术切除的髓母细胞瘤的原代外植体。对培养物进行免疫组织化学评估,检测一系列与神经元或胶质细胞分化相关的抗原决定簇。在所有培养物中均显示出III类β-微管蛋白、微管相关蛋白2(MAP2)以及程度较轻的tau蛋白的局灶性免疫定位。通过免疫印迹法在D283 Med细胞的明胶海绵基质培养物、单层培养物和悬浮培养物中也检测到了III类β-微管蛋白亚型、MAP2和tau蛋白。神经丝蛋白表位的染色高度可变,即使在源自同一原发肿瘤的不同培养物中也是如此,但神经丝蛋白随时间的变化(可能反映神经元分化)并未始终呈现。在所有培养物中均可见广泛的γ-烯醇化酶和局灶性突触素反应性,但未检测到S抗原染色。在一半的D283 Med细胞培养物中可变地存在Leu 7标记,但在源自5个原发肿瘤中4个的外植体中更为丰富。在所有时间点的D283 Med培养物中均始终发现波形蛋白。在D283 Med细胞系中未检测到胶质纤维酸性蛋白的免疫反应性。相反,在所有5例髓母细胞瘤手术标本中的散在星形细胞中均显示出该蛋白的染色。随着培养时间的增加,3例原发肿瘤在明胶海绵系统中培养时显示出胶质纤维酸性蛋白阳性细胞数量增加,但另外2例肿瘤的星形细胞成分极少。(摘要截短于250字)