Zusammenfassung
Bei 11 Patienten mit metastasierendem Melanom versuchten wir tumorinfiltrierende Lymphozyten (TIL) aus Metastasen (10 Lymphknotenmetastasen, 1 pulmonale Metastase) herzustellen, sie zu charakterisieren und ihre Zytotoxizität zu bestimmen. Die Resektate wurden mechanisch zerkleinert und anschließend durch ein Analysensieb (100 µm) gerieben. Die Zellsuspension wurde zur Hälfte über einen Ficollgradienten gereinigt zur anderen Hälfte belassen. Beide Fraktionen wurden mit und ohne Humanserum kultiviert, so daß 4 Kulturbedingungen untersucht wurden. Die Phänotypisierung der Zellen erfolgte mittels FACS-Analyse (Epics-Coulter), die Zytotoxizität wurde in einem 51 Chrom-Release-Assay gegen K 562-Zellen bestimmt. Eine Anzüchtung und Expansion von TIL gelang in 5 Fällen. Das beste Wachstum wurde bei ficollgereinigten, mit Humanserum kultivierten Zellen erreicht.
Summary
Tumor-infiltrating lymphocytes (TIL) were generated from 11 patients with metastatic malignant melanoma (10 lymph node metastases, 1 pulmonary metastasis). Tumor tissue was minced into fragments of 1 mm and then ground carefully through a sieve (100 µm). The cell suspension was divided. From one half, mononuclear cells were separated by Ficoll gradient centrifugation. Both fractions were cultured with and without human serum in the presence of interleukin-2. Expanded TIL were characterized by flow cytometry (Epics-Coulter), and cytotoxicity was determined by means of a standard 4 h chromium-51 release assay using K 562 cells as targets. Generation of TIL was successful in five of 11 cases. Lymph node metastases in malignant melanoma are suitable tissues for the culture and expansion of TIL. The best results were achieved with Ficoll-separated cells cultured with 1% human serum.
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© 1992 Springer-Verlag Berlin Heidelberg
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Wölfer, LU., Dummer, R., Becker, J.C., Taleghani, B.M., Burg, G. (1992). Herstellung Tumor-infiltrierender Lymphozyten aus Metastasen von Patienten mit malignen Melanomen. In: Burg, G., Hartmann, A.A., Konz, B. (eds) Onkologische Dermatologie. Fortschritte der operativen und onkologischen Dermatologie, vol 7. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-77690-8_7
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DOI: https://doi.org/10.1007/978-3-642-77690-8_7
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