Summary
Six rodent cell lines (36B10 rat glioma cells, 9L rat gliosarcoma cells, V79 Chinese hamster lung fibroblasts, EMT6/UW and EMT6/Ro mouse mammary sarcoma cells, and RIF-1 mouse fibrosarcoma cells) were tested for growth in cylindrical threads of Matrigel. These cells grew in the threads with doubling times of 17–23 h, reaching maximum cell densities on the order of 108 cells/ml. Histological sections of these threads showed a heterogeneous cell distribution: cells grew to confluence at the thread surface and at somewhat lower cell densities in the thread core. [H-3]thymidine labeling index and radiation sensitivity were measured for 9L and EMT6/UW cells in Matrigel threads. For both cell types, the labeling index in Matrigel was lower than observed in cell monolayers, with higher labeling indexes at the thread periphery than in the thread core. When these threads were grown in stirred medium, lower thread diameters, higher cell yields per thread, and higher labeling indices were obtained. EMT6 cell monolayers coated with Matrigel were less radiosensitive than cells in uncoated monolayers. This protective effect was eliminated by irradiating in the presence of 1 mg/ml misonidazole. EMT6 cells consume nearly three times as much oxygen (mole/cm3-sec) as do 9L cells, which are equally radiosensitive in monolayers with or without a Matrigel coating. The radiation sensitivity of EMT6/UW cells in Matrigel threads was similar to that for monolayers of plateau phase cells, whereas for 9L cells, the response in threads was more similar to exponentially growing cells. We conclude that Matrigel threads provide an alternativein vitro model for studying the radiation response of cells in a three-dimensional geometry.
Similar content being viewed by others
References
Casciari, J. J.; Sotirchos, S. V.; Sutherland, R. M. Glucose diffusivity in multicellular tumor spheroids. Cancer Res. 48:3095–3099; 1988.
Casciari, J. J.; Sotirchos, S. V.; Sutherland, R. M. Mathematical modeling of microenvironment and growth in EMT6/Ro multicellular tumor spheroids. Cell Proliferation 25:1–22; 1992a.
Casciari, J. J.; Sotirchos, S. V.; Sutherland, R. M. Variations in tumor cell growth rates and metabolism with oxygen concentration, glucose concentration and extracellular pH. J. Cell. Physiol. 151:386–394; 1992b.
Chou, J. L.; Shen, Z. X.; Stolfi, R. L., et al. Effects of extracellular matrix on the growth and casein gene expression of primary mouse mammary tumor cellsin vitro. Cancer Res. 49:5371–5376; 1989.
Daly, P. F.; Lyon, R. C.; Straka, E. J., et al.31P-NMR spectroscopy of human cancer cells proliferating in a basement membrane gel. FASEB J. 2:2596–2604; 1988.
Deen, D. F.; Hoshino, T.; Williams, M. E., et al. Development of a 9L multicellular spheroid system and its responses to BCNU and radiation. J. Natl. Cancer Inst. 64:1373–1382; 1980.
Foxall, D. L.; Cohen, J. S.; Mitchell, J. B. Continuous perfusion of mammalian cells embedded in agarose gel threads. Exp. Cell Res. 154:521–529; 1984.
Freyer, J. P.; Fink, N. H.; Schor, P. L., et al. A system for viably maintaining a stirred suspension of multicellular spheroids during NMR spectroscopy. NMR Biomed. 3:195–205; 1990.
Freyer, J. P.; Sutherland, R. M. Selective dissociation and characterization of cells from different regions of multicell tumor spheroids. Cancer Res. 40:3956–3965; 1980.
Gillies, R. J.; Galons, J. P.; McGovern, K. A., et al. Design and application of NMR-compatible bioreactor circuits for extended perfusion of high-density mammalian cell cultures. NMR Biomed. 6:95–104; 1993.
Grant, D. S.; Tashiro, K.; Segui-Real, B., et al. Two different laminin domains mediate the differentiation of human endothelial cells into capillary-like structures. Cell 58:933–943; 1989.
Hlatky, L.; Alpen, E. L.; Yee, M. K. Differences in the x-ray sensitivity of cells in different regions of the sandwich, a diffusion limited system for cell growth. Radiat. Res. 108:61–73; 1986.
Hohn, H. P.; Parker, C. R.; Boots, L. R., et al. Modulation of differentiation markers in human choriocarcinoma cells by extracellular matrix: on the role of a three dimensional matrix structure. Differentiation 51:51–70; 1992.
Kibbey, M. C.; Royce, L. S.; Dym, M., et al. Glandular-like morphogenesis of the human submandibular tumor cell line A253 on basement membrane components. Exp. Cell. Res. 198:343–351; 1992.
Kleinman, H. K.; McGarvey, M. L.; Liotta, L. A., et al. Isolation and characterization of type IV procollage, laminin, and heparan sulfate proteoglycan from the EHS sarcoma. Biochemistry 21:6188–6193; 1982.
Livesey, J. C.; Golden, R. N.; Shankland, E. G., et al. Measurement of tissue oxidation-reduction state with carbon-13 nuclear magnetic resonance spectroscopy. Cancer Res. 49:1937–1940; 1989.
Luk, C. K.; Keng, P. C.; Sutherland, R. M. Regrowth and radiation sensitivity of quiescent cells isolated from EMT6/Ro fed plateau monolayers. Cancer Res. 45:1020–1025; 1985.
Lyon, R. C.; Faustino, P. J.; Cohen, J. S. A perfusion technique for 13C NMR studies of the metabolism of 13C-labeled substrates by mammalian cells. Magn. Reson. Med. 3:663–672; 1986.
McGuire, P. G.; Seeds, N. W. The interaction of plasminogen activator with a reconstituted basement membrane matrix and extracellular macromolecules produced by cultured epithelial cells. J. Cell. Biochem. 40:215–227; 1989.
Melchiori, A.; Carlone, S.; Allavena, G., et al. Invasiveness and chemotactic activity of oncogene transformed NIH/3T3 cells. Anticancer Res. 10:37–44; 1990.
Mendonca, M. S.; Rodrigues, A.; Alpen, E. L. Quiescence in 9L cells and correlation with radiosensitivity and PLD repair. Radiat. Res. 117:433–447; 1989.
Mueller-Klieser, W.; Sutherland, R. M. Oxygen tensions in multicell spheroids of two cell lines. Br. J. Cancer 45:256–263; 1982.
Noel, A. C.; Calle, A.; Emonard, H. P., et al. Invasion of reconstituted basement membrane matrix is not correlated to the malignant metastatic cell phenotype. Cancer Res. 51:405–414; 1991.
Parodi, S.; Conio, M.; Munizzi, F., et al. Growth of cells from esophageal squamous cell carcinoma biopsies in a reconstituted basement membrane. Boll. Soc. Ital. Biol. Sper. 65:775–781; 1989.
Rannels, S. R.; Grove, R. N.; Rannels, D. E. Matrix-derived soluble components influence type II pneumocytes in primary culture. Am. J. Physiol. 256:C621–629; 1989.
Rasey, J. S.; Nelson, N. J. Response of anin vivo-in vitro tumour to x-rays and cytotoxic drugs: effect of tumour disaggregation method on cell survival. Br. J. Cancer 41, Suppl. IV:217–221; 1980.
Rasey, J. S.; Nelson, N. J. Repair of potentially lethal damage following irradiation with x rays or cyclotron neutrons: response of the EMT6/UW tumor system treated under various growth conditionsin vitro andin vivo. Radiat. Res. 85:69–84; 1981.
Reich, R.; Thompson, E. W.; Iwamoto, Y., et al. Effects of inhibitors of plasminogen activator, serine proteases, and collagenase IV on the invasion of basement membranes by metastatic cells. Cancer Res. 48:3307–3312; 1988.
Rofstad, E. K.; Sutherland, R. M. Radiation sensitivity of human ovarian carcinoma cell linesin vitro: effects of growth factors and hormones, basement membrane, and intercellular contact. Int. J. Radiat. Oncol. Biol. Phys. 15:921–929; 1988.
Schuetz, E. G.; Li, D.; Omiecinski, C. J., et al. Regulation of gene expression in adult rat hepatocytes cultured on a basement membrane matrix. J. Cell. Physiol. 134:309–323; 1988.
Steen, R. G. Response of solid tumors to chemotherapy monitored byin vivo 31P nuclear magnetic resonance spectroscopy: a review. Cancer Res. 49:4075–4085; 1989.
Toda, K.; Tsujioka, K.; Maruguchi, Y., et al. Establishment and characterization of a tumorigenic murine vascular endothelial cell line (F-2). Cancer Res. 50:5526–5530; 1990.
Topley, P.; Jenkins, D. C.; Jessup, E. A., et al. Effect of reconstituted basement membrane components on the growth of a panel of human tumor cell lines in nude mice. Br. J. Cancer 67:953–958; 1993.
Ugurbil, K.; Guernsey, D. L.; Brown, T. R., et al.31P NMR studies of intact anchorage-dependent mouse embryo fibroblasts. Proc. Natl. Acad. Sci. USA 78:4843–4847; 1981.
Uludag, H.; Sefton, M. V. Microencapsulated human hepatoma (HepG2) cells:in vitro growth and protein release. J. Biomed. Mater. Res. 27:1213–1224; 1993.
Vijayakumar, S.; Czerski, L.; Majors, A., et al. Phosphorous metabolite and cell cycle kinetic response of two human squamous cell carcinomas to radiation. Cancer Res. 52:5299–5306; 1992.
Wehrle, J. P.; Li, S. J.; Rajan, S. S., et al.31P and1H NMR spectroscopy of tumorsin vivo: untreated growth and response to chemotherapy. Ann. NY Acad. Sci. 508:200–215; 1987.
Williams, D. F.; Burke, J. M. Modulation of growth in retina-derived cells by extracellular matrices. Invest. Ophthalmol. & Visual Sci. 31:1717–1723; 1990.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Casciari, J.J., Chin, L.K., Livesey, J.C. et al. Growth rate, labeling index, and radiation survival of cells grown in the matrigel threadin vitro tumor model. In Vitro Cell Dev Biol - Animal 31, 582–589 (1995). https://doi.org/10.1007/BF02634310
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02634310