Cultured human neurocytoma cells from two neurosurgical patients were analysed for their immunocytochemical staining patterns and growth characteristics. In both cases, the cells stained positive for glial acidic fibrillary protein (GFAP) within one day of tissue culture in medium, with and without fetal calf serum, whereas the histological tumor specimens were negative. Both cases contained cells concomitantly expressing GFAP and synaptophysin (SNP) in the primary cultures. Epidermal growth factor (EGF) was mitogenic for the cultured cells but not platelet derived growth factor alpha (PDGF AA) or nerve growth factor (NGF). It is concluded that the human neurocytomas may represent neoplasms of a pluripotent neuroglial cell which can provide an interesting model to study the determinants for human glial/neuronal differentiation in vitro.
The immunocytochemical staining patterns of cultured glioma cells were investigated. Fifty nine individual cases were stained at different in vitro ages for glial fibrillary acidic protein, fibronectin, galactocerebroside, HNK-1/Leu 7, A2B5, vimentin, factor VIII and A4. Histologically, the cases were composed of eight low-grade astrocytomas, 11 high-grade astrocytomas, four low-grade oligodendrogliomas, seven high-grade oligodendrogliomas and 29 glioblastomas. The 45 cases were analysed within the first 3 weeks of culture, many of them as primary cultures. In 11 cases stainings were performed repeatedly at intervals of up to 6 months. Glial fibrillary acidic protein staining was positive in most of the early cultures of astrocytomas (low and high grade) and glioblastomas; expression in more than 50% of the cells was found in 1 of 5 low-grade astrocytomas, 5 of 11 high-grade astrocytomas and 14 of 29 glioblastomas. Two of the high-grade astrocytomas were stained once more after 6 weeks in culture and were found to be only 1% positive for glial fibrillary acidic protein but strongly positive for fibronectin. The same was true for five of the glioblastoma cases. Two of these cases remained glial fibrillary acid protein positive and developed into stable permanent cell lines. Only one case started with 1% of glial fibrillary acidic protein positive cells and later developed into a 99% glial fibrillary acidic protein positive cell line. Neither HNK-1/Leu 7 expression nor A2B5 staining appeared to have a relationship to the glial fibrillary acidic protein staining. It was observed that glial fibrillary acidic protein and HNK-1/Leu 7 were both 100% in some cases but that later one of the two antigens disappeared but not the other. The amount of glial fibrillary acidic protein staining does not allow the prediction of A2B5 staining. The study shows that initiation of primary cultures on an extracellular matrix yields more glial fibrillary acidic protein positive cells in primary cultures than have been found in other studies. It is concluded that only a rigid standardization of culture conditions will ensure the validity of comparisons of in vitro data obtained in primary cultures.
Cell culture has become an integral part of the daily routine of most oncology laboratories. It has enabled researchers to investigate a wide range of cellular parameters in a defined system in which the experimental conditions can be controlled and repeated. Although the manufacturers of tissue culture materials are continually improving their products, cell attachment and initial survival of primary cultures from tumor cells are still problems in many laboratories. Many different approaches have been taken to circumvent this problem, and coating of tissue culture dishes with attachment enhancers, such as polyamino acids (1), fibronectin (2), laminin (3), and collagen (4), has been found helpful. For a long time, it was known that endothelial cells produce a basement membrane, and this led to the use of bovine corneal basement membrane by Gospodarowicz et al. (5,6), in their research into the phenomenon of regeneration and nonregeneration of corneal endothelium in different species. The application of this bovine corneal extracellular matrix (bECM) has since been greatly expanded (5,6). bECM has found broad approval, and has been used for mammary carcinoma (7), urological tumors (8), and different kinds of pituitary adenomas (9,10) as well as CNS tumors (11).
No abstract
Cell culture is one of the major tools of cell biologists. It has also become an integral part of the daily routine of most oncology laboratories for the purpose of karyotyping, chemoresistance testing, or basic research. It provides investigators with an opportunity to investigate many cellular parameters and interactions in an in vitro system in which the experimental conditions can be controlled and repeated With many tissues, either human or animal, the problems of cell culture are cell attachment and initial survival. Particularly the primary cultures derived from tumor specimens are a problem in many laboratories. Apart from modifications in the composition of tissue-culture plastic materials, other approaches have been used to get around this problem, such as coating of tissue-culture dishes with attachment enhancers, such as polyamino acids (1), fibronectin (2), laminin (3), and collagen (4) Since it was known that endothelial cells are capable of producing a basement membrane even in vitro, bovine cornea1 endothelial basement membrane was explored by Gospodarowicz et al. for its role in regeneration and nonregeneration of cornea1 endothelium in different species This bovine cornea1 extracellular matrix (bECM) was found useful in the cell culture of a wide range of different cells (5, 6), and bECM as well as other ECMs were employed in the cell biology of tumor cells derived from mammary carcinomas (7), urological tumors (8), and different kinds of pituitary adenomas (9, 10), as well as CNS tumors (11), which is the topic of this chapter.
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