Summary
The establishment of a new glioma cell line, DBTRG-05MG, in a modified RPMI 1640 medium is described. The cells were derived from an adult female with glioblastoma multiforme who had been treated with local brain irradiation and multidrug chemotherapy; the tumor showed substantial change in histologic appearance compared to the original biopsy 13 mo. previously. The line has been successfully cryopreserved and passaged up to 20 times. The karyotype of the cells demonstrated it as a hypotetraploid line; the DNA index of 1.9 confirmed the karyotype analyses. By immunocytochemical analysis, the cell line reacted with polyclonal antibodies to vimentin, S100, and neuron specific enolase, reflecting its primitive neuroectodermal character. Positive immunostaining for epidermal growth factor receptor correlated with the excess of chromosome 7 seen in the karyotype. The cell line reacted negatively to antibodies against platelet-derived growth factor and its receptor, neuronal cell adhesion molecule, and glial fibrillary acidic protein. By flow cytometry, the cells were major histocompatibility class I antigen positive and class II antigen negative. Growth kinetic studies demonstrated an approximate population doubling time of 34 to 41 h and a colony forming efficiency of 71.4%. Western blot analysis showed the presence of low levels of normal-sized retinoblastoma protein. When compared to the patient’s lymphocyte DNA, no loss of heterozygosity of the p53 tumor suppressor gene was observed in the DBTRG-05MG cell line DNA.
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Batanian, J. R.; Ledbetter, S. A.; Wolff, R. K., et al. Rapid diagnosis of Miller-Dieker syndrome and isolated lissencephaly sequence by the polymerase chain reaction. Hum. Genet. 85:555–559; 1990.
Bigner, D. D.; Bigner, S. H.; Ponten, J., et al. Heterogeneity of genotypic and phenotypic characteristics of fifteen permanent cell lines derived from human gliomas. J. Neuropathol. Exp. Neurol. 40:201–229; 1981.
Bigner, S. H.; Bullard, D. E.; Pegram, C. N., et al. Relationship of in vitro morphologic and growth characteristics of established human glioma-derived cell lines to their tumorigenicity in athymic nude mice. J. Neuropathol. Exp. Neurol. 40:390–409; 1981a.
Bongcam-Rudloff, E.; Nister, M.; Betsholtz, C., et al. Human glial fibrillary acidic protein: complementary DNA cloning, chromosome localization, and messenger RNA expression in human glioma cell lines of various phenotypes. Cancer Res. 51:1553–1560; 1991.
Cordell, J. L.; Falini, B.; Erber, W. N., et al. Immunoenzymatic labeling of monoclonal antibodies using immune complexes of alkaline phosphatase and monoclonal anti-alkaline phosphatase (APAAP Complexes). J. Histochem. Cytochem. 32:219–229; 1984.
DeMuralt, B.; DeTribolet, N.; Diserens, A.-C., et al. Phenotyping of 60 cultured human gliomas and 34 other neuroectodermal tumors by means of monoclonal antibodies against glioma, melanoma and HLA-DR antigens. Eur. J. Cancer Oncol. 21:207–216; 1985.
Eng, L. F. Glial fibrillary acidic protein (GFAP): the major protein of glial intermediate filaments in differentiated astrocytes. J. Neuroimmunol. 8:203–214; 1985.
Green, M. A. When the products of oncogenes and anti-oncogenes meet. Cell 56:1–3; 1989.
Henn, W.; Blin, N.; Zang, K. D. Polysomy of chromosome 7 is correlated with overexpression of the erbB oncogene in human glioblastoma cell lines. Hum. Genet. 74:104–106; 1986.
Horn, G. T.; Richards, B.; Klinger, K. K. Amplification of a highly polymorphic VNTR segment by the polymerase chain reaction. Nucleic Acid Res. 17:2140; 1989.
Koprowski, H.; Herlyn, M.; Balaban, G., et al. Expression of the receptor for epidermal growth factor correlates with increased dosage of chromosome 7 in malignant melanoma. Somat. Cell Mol. Genet. 11:297–302; 1985.
Krishan, A.; Paika, K.; Frei, E. Cytofluorometric studies on the action of podophyllotoxin and epipodophyllotoxins (VM-26, VP-16-213) on the cell cycle traverse of human lymphoblasts. J. Cell Biol. 66:521–530; 1975.
Kruse, C. A.; Mitchell, D. H.; Lillehei, K. O., et al. Differences in two preparations of Interleukin-2 activated lymphocytes generated in vitro from peripheral blood of patients with malignant brain tumors. Cancer 64:1629–1637; 1989.
Lillehei, K. O.; Mitchell, D. H.; Johnson, S. D., et al. Long-term follow-up of patients with recurrent malignant gliomas treated with adjuvant adoptive immunotherapy. Neurosurgery 28:16–23; 1991.
Maxwell, M.; Naber, S. P.; Wolfe, H. J., et al. Expression of angiogenic growth factor genes in primary human astrocytomas may contribute to their growth and progression. Cancer Res. 51:1345–1351; 1991.
Murphee, A. L.; Benedict, W. F. Retinoblastoma: clues to human oncogenesis. Science 223:1028–1033; 1984.
Nister, M.; Libermann, T. A.; Betsholtz, C., et al. Expression of messenger RNAs for platelet-derived growth factor and transforming growth factor-a and their receptors in human malignant glioma cell lines. Cancer Res. 48:3910–3918; 1988.
Ogden, R. C.; Adams, D. A. Electrophoresis in agarose and acrylamide gels. In: Berger, S. L.; Kimmel, A. R., eds. Guide to molecular cloning techniques. Methods in Enzymology, vol. 152. San Diego: Academic Press; 1987:61–87.
Rowe, J. M.; Hemperly, J. J.; Sima, A. A. F., et al. Localization of neural cell adhesion molecules in human brain tumors. Am. Assoc. Neuropathol. 233A:366; 1991.
Schold, S. C., Jr.; Bullard, D. E.; Bigner, S. H., et al. Growth, morphology, and serial transplantation of anaplastic human gliomas in athymic mice. J. Neuro-Oncol. 1:5–14; 1983.
Shapiro, J. R.; Shapiro, W. R. Therapy modifies cellular heterogeneity in human malignant gliomas. Adv. Oncol. 8:21–29; 1992.
Steck, P. A.; Gallick, G. E.; Maxwell, S. A., et al. Expression of epidermal growth factor receptor and associated glycoprotein on cultured human brain tumor cells. J. Cell. Biochem. 32:1–10; 1986.
Studer, A.; deTribolet, N.; Diserens, A. C., et al. Characterization of four human malignant glioma cell lines. Acta Neuropathol. (Berlin) 66:208–217; 1985.
Takeshita, I.; Takaki, T.; Kuramitsu, M., et al. Characteristics of an established human glioma cell line, KNS-42. Neurol. Med. Chir. (Tokyo) 27:581–587; 1987.
Xu, H.-J.; Hu, S.-X.; Benedict, W. F. Lack of nuclear RB protein staining in Go/middle G1 cells: correlation to changes in total RB protein level. Oncogene 6:1139–1146; 1991.
Xu, H.-J.; Hu, S.-X.; Cagle, P. T., et al. Absence of retinoblastoma protein expression in primary non-small cell lung carcinomas. Cancer Res. 51:2735–2739; 1991a.
Zeltzer, P. M.; Schneider, S. L.; Marangos, P. J., et al. Differential expression of neural isozymes by human medulloblastomas and gliomas and neuroectodermal cell lines. JNCI 77:625–631; 1986.
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Kruse, C.A., Mitchell, D.H., Kleinschmidt-DeMasters, B.K. et al. Characterization of a continuous human glioma cell line DBTRG-05MG: growth kinetics, karyotype, receptor expression, and tumor suppressor gene analyses. In Vitro Cell Dev Biol - Animal 28, 609–614 (1992). https://doi.org/10.1007/BF02631035
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DOI: https://doi.org/10.1007/BF02631035