Cell membrane labeling with fluorescent dyes for the demonstration of cytokine-induced fusion between monocytes and tumor cells.
Fusion between monocytes and tumor cells has been suggested as a cause for tumor metastasis. The aim of the present study was to establish an in vitro fusion model representing the in vivo situation as close as possible. For this purpose fusion between cells was induced by cytokine containing conditioned medium. In order to prove that hybrid formation between tumor cells and monocytes occurs, a two-color-fusion-assay based on membrane labeling with the fluorochromes PKH 2 (green) and PKH 26 (red) was established. These fusion experiments were analyzed by microscopy and, in addition, by flow cytometry. The attempt to induce fusion between monocytes and several tumor cell lines of hematopoietic origin revealed quite diverse results. The most extensive hybrid formations were seen with TALL, a T-lymphocytic tumor line. The monocytic tumor line HL60 and the B-lymphocytic tumor line BL41 also clearly yielded hybrids with monocytes but in smaller numbers. With some other hematopoietic tumor lines no evidence for hybrid formation was detected. These studies indicate that fusion of normal monocytes with certain tumor cells may be induced under conditions that may occur in comparable manner in vivo.
Spötl
L
L
Institut für Hygiene, Leopold-Franzens-Universität Innsbruck, Austria.
Sarti
A
A
Dierich
M P
MP
Möst
J
J
D000328:Adult; D002232:Carbocyanines; D002459:Cell Fusion / Q000187:drug effects* / Q000502:physiology; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology / Q000648:ultrastructure; D016207:Cytokines / Q000494:pharmacology*; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D018922:HL-60 Cells; D006801:Humans; D008214:Lymphocytes / Q000502:physiology; D008565:Membrane Proteins / Q000032:analysis* / Q000502:physiology; D008853:Microscopy / Q000379:methods; D009000:Monocytes / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D009930:Organic Chemicals; D012235:Rhodamines; D014407:Tumor Cells, Cultured
D002232:Carbocyanines; D016207:Cytokines; D005456:Fluorescent Dyes; D008565:Membrane Proteins; D009930:Organic Chemicals; C070080:PKH 26; C068241:PKH-2; D012235:Rhodamines; C024286:3,3'-dioctadecylindocarbocyanine; C009434:tetramethylrhodamine isothiocyanate; D016650:Fluorescein-5-isothiocyanate