Morphological Heterogeneity and Phenotype Modifications during Long Term in Vitro Cultures of Six New Human Glioblastoma Cell Lines (original) (raw)
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The American journal of pathology, 1987
Explants derived from human gliomas have been characterized with respect to their cellular outgrowth pattern after 1-22 weeks in culture. A mat of cells which were fibronectin (FN)-positive and glial fibrillary acidic protein (GFAP)-negative (hereafter designated FN+ cells) with a polygonal, flat morphology covered the growth substrate in a swirling pattern for a mean diameter of 9.2 mm around FN+ explants. FN+ cells showed ruffled plasmalemma, dilated rough endoplasmic reticulin (RDR), and extracellular filamentous strands. Rare desmosomes were compatible with at most minor leptomeningeal components or differentiation. FN+ cells predominated in six of seven cultures at passage 2, and their features were the same from various high-grade gliomas and gliosarcoma. Around other explants, elongated or stellate cells which were GFAP+ and FN- grew in a netlike pattern with little cell-to-cell contact. These GFAP+ cells surrounded explants at a mean diameter of 2 mm, substantially less than...
Vojnosanitetski pregled, 2014
Background/Aim. The cell line C6 is a continuous cell line of rat glioma and, as a transplantable line, is frequently used for induction into in vivo model of primary brain tumor. It is believed that, pursuant to its histological traits and biological behavior, this experimental tumor corresponds to human anaplastic astrocytoma of grade II/III, which is characterized by proliferative and invasive potency, and marked cell differentiation. The aim of this study was to determine macroscopic analysis of rat brain with implanted tumor during tumorigenesis, histological features of tumor cells of induced brain tumor and markers of proliferation (proliferation cell nuclear antigen - PCNA, cytokeratin - CK 19) and differentiation (glial fibrillary acidic protein -GFAP) in rat brain with implanted tumor. Methods. To determine histological structure of the brain with implanted C6 cells, we used brain sections stained for hematoxylin-eosin or kresyl violet, whereas other sections were immunohi...
Characteristics of an established human glioma cell line, KNS-42
Neurologia medico-chirurgica, 1987
We established a human glioma cell line derived from a malignant glioma and evaluated it by im munochemical techniques and antibodies to astroglial (glial fibrillary acidic protein: GFAP, S-100 protein), oligodendroglial (myelin basic protein, galactocerebroside), neuronal (neuron-specific eno lase: NSE, neurofilament triplet proteins), and mesenchymal (vimentin, fibronectin) antigens. The cell line was epithelial in sparse culture and glial in dense culture. GFAP was expressed by most cells in sparse culture and primarily by overlying cells in dense culture. The amount of GFAP depended on the cell density and ranged from 530 to 990 ng/mg of protein. The cells contained much more vimentin than GFAP. Neurofilament proteins, S-100 protein, myelin basic protein, and galactocerebroside were undetectable. The cells synthesized a small amount of fibronectin and released it into the medium. The amount of NSE was unrelated to cell density, ranged from 13 to 19 ng/mg of protein, and was present in greater quantity in cells cultured in a medium containing 1 ul/ml of sodium lactate. We conclude that KNS-42 is a permanent astrocytic glioma cell line; it expresses GFAP, NSE, vimen tin, and fibronectin, and is morphologically and functionally dependent on its environment.
Characterization of an established human malignant glioma cell line: LN-18
Acta Neuropathologica, 1981
A human malignant glioma cell line, has been established in monolayer culture and subcultured for more than 115 passages. LN-18 cells grow in vitro as bipolar or stellate cells with pleomorphic nuclei, have a doubling time of about 72 h and a plating efficiency of 3 %. The glial nature of these cells has been assessed by ultrastructural examination. The synthesis of glial fibrillary acidic and S-100 proteins could not be demonstrated, although the initial biopsy tissue and the early cultures were positive for the former. The presence of [a-like antigens on the surface of these cells was demonstrated using allo and xeno antisera. LN-18 cells were also shown to synthesize large quantities of fibronectin. The injection of LN-18 cells into nude mice induced the formation of solid tumor masses that could be retransplanted every 3 weeks and showed a morphology comparable to that of the initial biopsy. Karyotype analysis revealed the presence of three marker chromosomes, constantly present before and after hetero-transplantation.
Establishment and Characterization of a Human Glioblastoma Multiforme Cell Line
Cancer Genetics and Cytogenetics, 1998
Cell lines provide a useful system for further understanding the biology of glioblastoma multiforme. In this study, a new glioblastoma multiforme cell line, GATAGM-96 (Gulhane Askeri Tip Akademisi-Glioblastoma Multiforme-96), was established from a tumor specimen removed from an 80year-old male patient who underwent surgery for intracranial tumor. Morphologic examination, immunocytochemical staining, growth kinetics, and karyotypic characteristics of this cell line were studied. The cytoskeleton was positive for neuron-specific enolase, vimentin, and neurofilament, and it was negative for glial fibrillary acidic protein, S-100 protein, p53 protein, epidermal growth factor, and transforming growth factor ␣ . Growth kinetic studies demonstrated an approximate population doubling time of 38 to 42 h and a colony forming efficiency of 83.3%. The karyotype of the cells demonstrated it as hyperdiploid, with a large subpopulation of polyploid cells. There were numerous structural and numerical chromosome aberrations; most of them were present as clonal events. The phenotypic and chromosomal features detailed on the GATAGM-96 cell line should make it a useful addition to the cell lines currently available for in vitro and in vivo studies of glioblastoma multiforme. © Elsevier Science Inc.
Morphological characterization of a human glioma cell l ine
Cancer cell international, 2005
A human malignant continuous cell line, named NG97, was recently established in our laboratory. This cell line has been serially subcultured over 100 times in standard culture media presenting no sign of cell senescence. The NG97 cell line has a doubling time of about 24 h. Immunocytochemical analysis of glial markers demonstrated that cells are positive for glial fibrillary acidic protein (GFAP) and S-100 protein, and negative for vimentin. Under phase-contrast microscope, cultures of NG97 showed cells with variable morphological features, such as small rounded cells, fusiform cells (fibroblastic-like cells), and dendritic-like cells. However, at confluence just small rounded and fusiform cells can be observed. At scanning electron microscopy (SEM) small rounded cells showed heterogeneous microextentions, including blebs and filopodia. Dendritic-like cells were flat and presented extensive prolongations, making several contacts with small rounded cells, while fusiform cells present...
Immunocytochemical studies using polyclonal antibodies to epidermal growth factor (EGF) and transforming growth factor (TGF) alpha and beta wereperformed on 20 cases ofhuman gliomas. EGF immunoreactive material was detected in both benign and malignant glial tumors. In addition, EGF immunoreactive material was detected in normal brain. TGF-beta was detected in both benign and malignant tumors, but was not detected in normal brain. In contrast, TGF-alpha was highly conserved in its expression, occurring predominantly in malignant compared with benign or normal brain tissue (P < 0.0001). In malignant gliomas, glioblastomas contained 76% TGF-alpha reactivity (immunoreactive product), and anaplastic types contained 85% reactivity. Benign gliomas contained only 13 % TGF-alpha reactivity. These findings support the role of TGF-alpha as an oncoprotein marker in brain neoplasms. (Am J Pathol 1989, 134:895-902) Transforming growth factors (TGF) are low molecular weight polypeptides that reversibly induce anchorage-independent growth of nontransformed anchorage-dependent cells. At least two types of transforming growth factors have been described-alpha and beta. TGF alpha is a M, 6000 single-chain polypeptide that shares 30 to 35% homology with human and rat epidermal growth factor (EGF).1 Both EGF and TGF-alpha can compete for the same membrane receptors.2 In contrast to EGF, TGF-alpha secretion is seen primarily in the transformation of cells to a malignant phenotype. The release of TGF-alpha by transformed cells is thought to be the result of oncogene activation rather than a consequence of a cellular change during the process of transformation.3 The link between oncogenes and growth factors has been established for many polypeptide growth factors.4 The autonomous production of growth factors has led to the autocrine mechanism for neoplastic conversion.
Journal of Neurosurgery, 1982
✓ This report presents the results of a study using multiple techniques of the established human cell line, LM, which has been developed in culture medium from a patient with a right temporoparietal glioblastoma. This cell line has human subtetraploid karyotype and has several features of a transformed line in culture. These include continuous propagation for 10 years, ability to form tumor nodules when transplanted into immunologically suppressed hamsters, and pleomorphic appearance. Ultrastructurally, it is characterized by multiple nuclei, few actin cables, and numerous surface-membrane microvilli, as well as abundant 9- to 10-nm cystoplasmic filaments. By its immunological reactivity, the line can be shown to contain glial fibrillary acidic protein at low levels, consistent with its glial origin and continued nature. Dibutyryl cyclic adenosine monophosphate (db-cAMP) induces formation of long astrocytic-like processes as well. Its membrane electrical characteristics include a lo...