[Automated cytofluorometry of solid malignant tumors after cell dispersal by means of intraperitoneal cultivation of cell complexes in young rats (author's transl)].
The use of isolated cells is a basic requirement besides data processing techniques to obtain optimum results with the automated flow cytofluorometry. Several techniques have been employed in order to get cell dispersal. But the claim for reproducible and representative cell samples cannot exactly be fulfilled: gross parts of cells are damaged by any dispersal method applied on solid tissues. Therefore we want to introduce a special method for cell isolation, which avoids mechanical and chemical damage of the treated cells. For this the intraperitoneal tolerance of growing tumor cells in young animals was used, in the species of which solid growth of such tumor cells is established. The investigations reported here were carried out with the Walker-tumor 256 on the rat as a model for solid tumor growth. Our interest was focused on the determination of cellular CNA distribution in tumor cells and contaminating somatic cells grown in the ascitic fluids.


METHODS
Walker-tumors are permanently cultivated on Sprague-Dawley rats. The solid tumors (1 cm diameter) were removed under sterile conditions, cut with scissors, and resuspended in 0.9% saline. Ten rats (37 days old, body weight 90-120 g) had received an intraperitoneal injection of 1 ml Liquemin (5000 Roche). One million tumor cells could be obtained by such a procedure. After spontaneous sedimentation of cells in order to reduce the number of erythrocytes, the cells were processed and fixed according to Trujillo and Van Dilla (1972), digested with heated RNase (1 mg/ml tris-buffer; 0.18 M and pH 7.5), pepsin treated, and then stained with ethidium bromide (40 mug/ml tris-buffer) for 30 min in the cold. After rinsing with distilled water several times the cells were resuspended in buffer and measured within an hour immediately after passing a 10 mum filter to avoid cell aggregates...
D000818:Animals; D002279:Carcinoma 256, Walker / Q000473:pathology*; D046508:Culture Techniques; D004273:DNA, Neoplasm / Q000032:analysis; D005470:Fluorometry / Q000379:methods*; D008297:Male; D008325:Mammary Neoplasms, Experimental / Q000473:pathology*; D009362:Neoplasm Metastasis; D010529:Peritoneal Cavity; D051381:Rats
D004273:DNA, Neoplasm
Automatische Zytofluorometrie solider Malignome nach Zellenvereinzelung durch intraperitoneale Anzucht von Zellkomplexen in jungen Ratten