Summary
In the present study we describe a new method to cultivate human tumors, which allows organoid differentiation under in vitro conditions. Diverse tumors of different origin and various histopathology which had been heterotransplanted to athymic mice were dissociated into single cells and seeded at high cell density onto a membrane filter consisting of cellulose nitrate at the gas-medium interface. Within a few days, the tumor cells reorganized and differentiated into organoid structures which exhibited the typical histological characteristics of the original tissues. Due to the formation of organoid aggregates, which was also previously seen with normal fetal cells, this type of culture has been described as ‘organoid culture’. In the case of adenocarcinomas of the lung and the colon including the rectum, glandular structures with central lumina, adjacent microvilli, and junctional complexes were formed. Numerous specific intercellular contacts such as desmosomes and tight junctions occurred as well as interdigitations of adjacent cell membranes. In a tumor of the rectum, a typical brush border differentiated at the surface of the reorganized tumor-tissue aggregate. Epidermoid carcinomas of the head and neck developed structures resembling the spinous layer of the epidermis, exhibiting numerous desmosomes and intracytoplasmic bundles of tonofilaments radiating into the desmosomes. Most tumors produced a fragmentary monolayered or multilayered basal lamina of similar morphological appearance as under in vivo conditions. These results illustrate the organoid reorganization and differentiation of human tumor cells under the experimentally rather simple conditions of the organoid culture systems and clearly demonstrate that this in vitro system comes close to the in vivo situation as far as certain differentiation phenomena are concerned.
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Bellet RE, Danna V, Mastrangelo MJ, Berd D (1979) Evaluation of a “nude” mouse-human tumor panel as a predictive secondary screen for cancer chemotherapeutic agents. J Natl Cancer Inst 63:1185–1188
Bertoncello I, Bradley TR (1987) Human cell cultures for screening anti-cancer drugs. Trends Pharmacol Sci 8:249–251
Bogden AE, Von Hoff DD (1984) Comparison of the human tumor cloning and subrenal capsule assays. Cancer Res 44:1087–1090
Braakhuis BJM, Sneeuwloper G, Snow GB (1984) The potential of the nude mouse xenograft model for the study of head and neck cancer. Arch Otorhinolarynol 239:69–79
Bulbul MA, Pavelic K, Slocum HK, Frankfurt OS, Rustum YM, Huben RP, Bernacki RJ (1986) Growth of human urologic tumors on extracellular matrix. J Urol 136:512–516
Christensen TG, Burke B, Dexter DL, Zamcheck N (1985) Ultrastructural evidence of dimethylformamide-induced differentiation of cultured human colon carcinoma cells. Increased expression of desmosomes. Cancer 56:1559–1565
Damjanov I, Damjanov N, Knowles BB, Engvall E (1985) Origin of laminin in the extracellular matrix of human tumor xenografts in nude mice. Virchows Arch [B] 49:45–52
David G, Van Den Berghe H (1983) Transformed mouse mammary epithelial cells synthesize undersulfated basement membrane proteoglycan. J Biol Chem 258:7338–7344
Fiebig HH, Schuchhardt C, Henss H, Fiedler L, Löhr GW (1984) Comparison of tumor response in nude mice and in the patients. Behring Inst Mitt 74:343–352
Freeman AE, Hoffman RM (1986) In vivo-like growth of human tumors in vitro. Proc Natl Acad Sci USA 83:2694–2698
Fricker SP, Buckley RG (1989) Characterisation of a human tumour cell line panel for screening of anti-tumour agents in vitro. Biochem Soc Trans 17:1049–1050
Hamburger A, Salmon SE (1977) Primary bioassay of human tumor stem cells. Science 197:461–463
Hoffman RM, Connors KM, Meerson-Monosov AZ, Herrera H, Price JH (1989) A general native-state method for determination of proliferation capacity of human normal and tumor tissues in vitro. Proc Natl Acad Sci USA 86:2013–2017
Honegger P, Richelson E (1977) Biochemical differentiation in aggregating cell cultures of different fetal brain regions. Brain Res 133:329–339
Ingber DE, Madri JA, Jamieson JD (1981) Role of basale lamina in neoplastic disorganization of tissue architecture. Proc Natl Acad Sci USA 78:3901–3905
Kangas L, Käpylä H, Roberts PJ (eds) (1985) Subrenal capsule assay. Ann Chir Gyn 74 [Suppl 199]:1–65
Kao JW-Y, Collins JL (1989) A rapid in vitro screening system for the identification and evaluation of anticancer drugs. Cancer Invest 7:303–311
Karst W, Merker HJ (1989) The differentiation behaviour of MDCK cells grown on matrix components and in collagen gels. Cell Differentiation 22:211–224
Köpf-Maier P (1986) Dying and regeneration of human tumor cells after heterotransplantation to athymic mice. Histol Histopath 1:383–390
Köpf-Maier P, Kestenbach U, Merker HJ (1991) Immunfluoreszenzmikroskopische Studien zum Aufbau und zur Entstehung der Basalmembran in heterotransplantierten menschlichen Tumoren. Verh Anat Ges 84 (in press)
Landry J, Freyer JP (1984) Regulatory mechanisms in spheroidal aggregates of normal and cancerous cells. In: Acker H, Carsson J, Durand R, Sutherland RM (eds) Speroids in cancer research: methods and perspectives, Springer, New York, pp 50–66
Landry J, Bernier D, Quellet C, Goyette R, Marceau N (1985) Speroidal aggregate culture of rat liver cells: bistotypic reorganization, biomatrix deposition, and maintenance of functional activities. J Cell Biol 101:914–923
Lawson AW, riches AC, Weaver JPA (1986) Human bladder tumors in organ culture. J Urol 135:1061–1065
Liotta LA, Kleinerman J, Catanzaro P, Rynbrandt D (1977) Degradation of basement membrane by murine tumor cells. J Natl Cancer Inst 58:1427–1431
Liotta LA, Terranova VP, Lanzer WL, Russo R, Siegel GP, Garbisa S (1982) Basement membrane attachment and degradation by metastatic tumor cells. In: Kühn K, Schöne H, Timpl R (eds) New trends in basement membrane research. Raven Press, New York, 277–285
Matsuoka H, Sugimachi K, Yano K (1987) Chemosensitivity testing for anticancer drugs using confluent monolayers. Med Sci Res 15:837–838
Merker HJ, Zimmermann B, Barrach HJ (1984) The significance of cell contacts for the differentiation of the skeletal blastema. Acta Biol Hung 35:195–203
Moscona A (1952) Cell suspension from organ rudiments of chick embryos. Exp Cell Res 3:535–539
Müller-Klieser W (1987) Multicellular spheroids. A review on cellular aggregates in cancer research. J Cancer Res Clin Oncol 113:101–122
Neubert D (1982) The use of culture techniques in studies of prenatal toxicity. Pharmac Ther 18:397–434
Neumeier R, Maurer HR, Maschler R, Bombik BM (1981) A micromethod to culture granulocyte colonies from human bone marrow in agar-containing glass capillaries. Exp Hematol 9:44–51
Ono J, Takaki R, Okano H, Fukuma M (1979) Long-term culture of pancreatic islet cells with special reference of the B-cell function. In Vitro 15:95–102
Povlsen CO, rygaard J (1971) Heterotransplantation of human adenocarcinomas of the colon and rectum to the mouse mutant nude. A study of nine consecutive transplantations. Acta Path Microbiol Scand (A) 79:159–169
Povlsen CO, Rygaard J (1972) Heterotransplantation of human epidermoid carcinomas to the mouse mutant nude. Acta Path Microbiol Scand (A) 80:713–717
Salmon SE (1984) Human tumor colony assay and chemosensitivity testing. Cancer Treat Rep 68:117–125
Salmon SE, Alberts DS, Meyskens FL, Duric BGM, Jones SE, Soehnlen B, Young L, Chen HSG, Moon TE (1980) Clinical correlations of in vitro drug sensitivity. In: Salmon SE (ed) Cloning of human tumor stem cells. Progr Clin Biol Res 48:223–245
Sutherland RM (1988) Cell and environmental interactions in tumor microregions: The multi-cell spheroid model. Science 240:177–184
Vladavsky I, Ariav Y, Atzmon R, Fuks Z (1982) Tumor cell attachment to the vascular endothelium and subsequent degradation of the subendothelial extracellular matrix. Exp Cell Res 140:149–159
Vogh J, Trempe G (1975) New human tumor cell lines. In: Fogh J (ed) Human tumor cells in vitro. Plenum Press, New York, 115–159
Von Hoff DD, Harries GJ, Johnson G, Glaubiger D (1980) Initial experience with the human tumor stem cell assay: potential and problems. In: Salmon SE (ed) Cloning of human tumor stem cells. Progr Clin Biol Res 48:113–124
Wolff E, Wolff E (1975) Current research with organ cultures of human tumors. In: Fogh J (ed) Human tumor cells in vitro. Plenum Press, New York, 207–240
Zimmermann B (1987) Lung organoid culture. Differentiation 36:86–109
Zimmermann B, Tsambaos D (1985) Evaluation of the sensitive step of inhibition of chrondrogenesis by retinoids in limb mesenchymal cells in vitro. Cell Differentiation 17:95–103
Zimmermann B, Wachtel HC, Somogyi H, Merker HJ, Bernimoulin IP (1988) Bone formation by rat calvarial cells grown at high density in organoid culture. Cell Differ Dev 25:145–154
Zimmermann B, Wachtel HC, Somogyi H (1990) Endochondral mineralization in cartilage organoid culture. Cell Differ Dev 31:11–22
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Köpf-Maier, P., Zimmermann, B. Organoid reorganization of human tumors under in vitro conditions. Cell Tissue Res 264, 563–576 (1991). https://doi.org/10.1007/BF00319046
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DOI: https://doi.org/10.1007/BF00319046