Summary
The Rotating-Wall Vessel (RWV) is a novel in vitro cell culture system used to successfully culture a cell line derived from a heterologous mixed mullerian tumor cell of the ovary. Although the original tumor was comprised of both epithelial and mesodermal components, long-term culture in conventional flasks established a cell line from this tumor with homogenous epitheliallike growth characteristics (1). Cells from Passage 36 were seeded into a Rotating-Wall Vessel containing Cytodex-3 microcarrier beads. Scanning electron micrographs of tumor cells cultured for 32 d in the RWV showed the presence of heterogeneous cell populations organized into three-dimensional tissuelike architecture. Immunocytochemical analysis confirmed the cellular heterogeneity, as demonstrated by expression of both epithelial and mesenchymal antigens. Reverse transcription polymerase chain reaction amplification demonstrated the presence of mRNA for cellular oncogenes HER-2/neu, H-ras, K-ras, and tumor suppressor p53. Thus, there are two advantages to propagation of tissue in the RWV culture system: (a) tissue diversification representing populations present in the original tumor, and (b) the three-dimensional freedom to organize tissues morphologically akin to those observed in vivo. These data indicate that the RWV culture system is suitable for generating large quantities of ovarian tumor cells in vitro that are amenable to immunocytochemical, oncogenic, morphologic characteristics demonstrated in vivo.
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Becker, J. L.; Finan, M.; Widen, R. H. Oncogene expression by a mullerian tumor cell line. In Vitro Cell. Dev. Biol. 28II:149A; 1992.
Becker, J. L.; Papenhausen, P. R.; Widen, R. H. Cytogenetic, morphologic, and oncogene analysis of cell line derived from heterologous mixed mullerian tumor of the ovary. In Vitro Cell. Dev. Biol. 33A:325–331, 1997.
Becker, J. L.; Prewett, T. L.; Spaulding, G. F., et al. Three-dimensional growth and differentiation of ovarian tumor cell line in High-Aspect Rotating-Wall Vessel: morphologic and embryologic consideration. J. Cell. Biochem. 57:283–286; 1993.
Bellet, R. E.; Danna, V.; Mastrangelo, M. J., et al. Evaluation of a nude mouse-human tumor panel as a predictive secondary screen for cancer chemotherapeutic agents. J. Natl. Cancer Inst. 63:1185–1188; 1979.
Crickard, K.; Crickard, U.; Yoonessi, M. Human ovarian carcinoma cells maintained on extracellular matrix versus plastic. Cancer Res. 43:2762–2767; 1983.
Crickard, K.; Crickard, U.; Yoonessi, M., et al. Extracellular matrixcoated culture dishes: an in vitro model for primary human gynecologic tumors. Cancer Genet. Cytogenet. 19:51–56; 1986.
Crickard, K.; Niedbala, M. J.; Crickard, U., et al. Characterization of human ovarian and endometrial carcinoma cell lines established on extracellular matrix. Gynecol. Oncol. 32:163–173; 1989.
Emoto, M.; Iwasaki, H.; Kikuchi, M., et al. Two cell lines established from mixed mullerian tumors of the uterus. Cancer 59:1759–1768; 1992.
Geisinger, K. R.; Dabbs, D. J.; Marshall, R. B. Malignant mixed mullerian tumors. An ultrastructural and immunohistochemical analysis with histogenetic considerations. Cancer 59:1781–1790; 1987.
Giavazzi, R.; Campbell, D. E.; Jessup, J. M., et al. Metastatic behavior of tumor cells isolated from primary and metastatic human colorectal carcinomas implanted into different sites in nude mice. Cancer Res. 46:1928–1933; 1986.
Giovanella, B. C.; Stehlin, J. S.; Williams, Jr., L. J., et al. Heterotransplantation of human cancers into nude mice: a model system for human cancer chemotherapy. Cancer 42:2269–2281; 1978.
Goodwin, T. J.; Jessup, J. M.; Wolf, D. A. Morphologic differentiation of colon carcinoma cell lines HT-29 and HT-29KM in rotating-wall vessels. In Vitro Cell. Dev. Biol. 28A:47–60; 1992.
Goodwin, T. J.; Prewett, T. J.; Spaulding, G. F., et al. A model for culture of primary breast carcinoma tissues in rotating-wall vessels (RWV). Mol. Biol. Cell 4S:15a; 1993.
Goodwin, T. J.; Prewett, T. L.; Wolf, D. A., et al. Reduced shear stress: a major component in the ability of mammalian tissues to form three-dimensional assemblies in simulated microgravity. J. Cell. Biochem. 51:301–311; 1993.
Goodwin, T. J.; Schroeder, W. F.; Wolf, D. A., et al. Rotating-wall vessel coculture of small intestine as a prelude to tissue modeling: aspects of simulated microgravity. Proc. Soc. Exp. Biol. Med. 202(2):181–192; 1993.
Kozlowski, J. M.; Fidler, I. J.; Campbell, D., et al. Metastatic behavior of human tumor cell lines grown in the nude mouse. Cancer Res. 44:3522–3529; 1984.
Krishan, A. A rapid flow cytofluorometric analysis of mammalian cell cycle by propidium iodide. J. Cell Biol. 66:188–193; 1975.
Masuda, A.; Takeda, A.; Fukami, H., et al. Characteristics of cell lines established from a mixed mesodermal tumor of the human ovary. Carcinomatous cells are changeable to sarcomatous cells. Cancer 60:2696–2703; 1987.
Niedbala, M. J.; Crickard, K.; Bernacki, R. J. Interactions of human ovarian tumor cells with human mesothelial cells grown on extracellular matrix. Exp. Cell Res. 160:499–513; 1985.
Niedbala, M. J.; Crickard, K.; Bernacki, R. J. In vitro degradation of extracellular matrix by human ovarian carcinoma cells. Clin. & Exp. Metastasis 5:181–197; 1987.
Noorduyn, L. A.; Herman, C. J. The relation between mixed mesodermal tumors and adenocarcinomas of the ovary. An immunopathologic study. Eur. J. Cancer & Clin. Oncol. 23:157–162; 1987.
Ovajera, A. A.; Houchena, D. P.; Barker, A. D. Chemotherapy of human tumor xenografts in genetically athymic mice. Ann. Clin. Lab. Sci. 8:50–56;1978.
Povlsen, C. O.; Jacobsen, G. K. Chemotherapy of a human malignant melanoma transplanted in the nude mouse. Cancer Res. 35:2790–2796; 1975.
Prewett, T. L.; Goodwin, T. J.; Spaulding, G. F. Three-dimensional modeling of T-24 human bladder carcinoma cell line: a new simulated microgravity culture vessel. J. Tiss. Cult. Meth. 15:29–36; 1993.
Puts, J. J.; Moesker, O.; Aldeweireldt, J., et al. Application of antibodies to intermediate filament proteins in simple and complex tumors of the female reproductive tract. Int. J. Gynecol. Pathol. 6:257–274; 1987.
Rubin, A. The histogenesis of carcinosarcoma (mixed mesodermal tumor) of the uterus as revealed by tissue culture studies. Am. J. Obstet. Gynecol. 269–274; 1959.
Schwarz, R. P.; Goodwin, T. J.; Wolf, D. A. Cell culture for three-dimensional modeling in rotating-wall vessels: an application of simulated microgravity. J. Tiss. Cult. Meth. 14:51–58; 1992.
Simon-Assmann, P.; Bouziges, F.; Vigny, M., et al. Origin and deposition of basement membrane heparin sulfate proteoglycan in the developing intestine. J. Cell Biol. 109:1837–1846; 1989.
Takeda, A.; Matsuyama, M.; Kuzuya, K., et al. Mixed mesodermal tumor of the ovary with carcinoembryonic antigen and alkaline phosphatase production. Cancer 53:103–112; 1984.
Taylor-Papadimitriou, J.; Berdichevsky, F.; D’Souza, B., et al. Human models of breast cancer. Cancer Surv. 16:59–78; 1993.
Tsao, Y. D.; Goodwin, T. J.; Wolf, D. A., et al. Responses of gravity level variations on the NASA/JSC bioreactor system. The Physiologist 35:S49-S50; 1992.
van Haaften-Day, C.; Russell, P.; Brammah-Carr, S. Two homologous mixed mullerian tumor lines of the ovary and their characteristics. Cancer 65:1753–1761; 1990.
Vescio, R. A.; Conners, K. M.; Kubota, T., et al. Correlation of histology and drug response of human tumors grown in native-state three-dimensional histoculture and nude mice. Proc. Natl. Acad. Sci. 88:5163–5166; 1991.
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Goodwin, T.J., Prewett, T.L., Spaulding, G.F. et al. Three-dimensional culture of a mixed mullerian tumor of the ovary: Expression of in vivo characteristics. In Vitro Cell.Dev.Biol.-Animal 33, 366–374 (1997). https://doi.org/10.1007/s11626-997-0007-4
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DOI: https://doi.org/10.1007/s11626-997-0007-4