TISSUE culture techniques are invaluable for the study of biological processes at the cellular level and particularly so in the experimental study of human tumours which cannot easily be maintained after removal from the patient. Although a number of cell lines have been established from human tumours (Moore and Koike, 1964) following the establishment of the HeLa cell line from a carcinoma of cervix by Gey et al. (1952), there are no readily available lines from human tumours of the urinary tract. Short term cultures of bladder tumours have been reported by Burrows et al. (1917), Bregman andBregman (1961) andWalker et al. (1965) among others. Jones (1967) reported one cell line from a carcinoma of bladder which had been maintained for 20 months. Although the original cultures contained " epithelial " and " fibroblastic " cells, the " epithelial " elements died out, and the established line was considered to be of normal connective tissue origin.In order to study the cytological behaviour of urothelial neoplasms in vitro and in vivo, and their response to cytotoxic agents, a number of tumours were selected for tissue culture in an attempt to establish permanent cell lines with the characteristics of the parent neoplastic transitional epithelium. MATERIAL AND METHODSCultures were prepared from cell suspensions or explants. Cell suspensions were prepared by mincing with crossed scalpels. The mince was then passed through a 20 gauge needle. In the more solid tumours, suspensions were prepared by trypsinization at 36 5°C., in fluted 250 ml. Erlenmeyer flasks. The trypsin (Tryptar, Almour) was used at a concentration of 500 units!

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ALTHOUGH no chromosome karyotypes specific to solid tumours have so far been reported, most human malignant tumours studied have had aneuploid chromosome complements (Spriggs, Boddington and Clarke, 1962; Yamada, Tagaki and Sandberg, 1966; Miles, 1967). The role of chromosomes in the malignant process is still uncertain, and changes gross enough to be detected by current methods may be a result rather than a cause (Hauschka, 1961;Sandberg, 1966). However such changes do characterise the progression of tumour growth, and hence are relevant to the study of malignancy. The techniques for direct karyotyping of solid tumours are slow, and for this reason the series reported are usually small. MATERIALS AND METHODSMetaphase chromosome counts and karyotypes were analysed in 10 testicular tumours. Tissue was obtained as fresh as possible by attendance in the operating theatre whenever possible. If the delay before receiving the tissue was greater than 30 minutes after excision there was noticeable deterioration in the quality of the preparations and in the number of suitable metaphases isolated. Small pieces of tissue, about 1 cm. in diameter, were chopped with scissors to give a fine suspension in 5 ml. of tissue culture medium 199 (Glaxo). This cell suspension was incubated at 370 C. for 1 hour after the addition of Colcemid (Ciba) 4 /tg./ml.The cells were then transferred to 0 95 % sodium citrate for 15 minutes before fixation in acetic-methanol 1: 3. Slides were prepared by air-drying (Rothfels and Siminovitch, 1958) and stained in 1 % lacto-aceto-orcein. Metaphases were selected and photographed by phase microscopy. Chromosomes were counted in 50 separate metaphases, and karyotypes were prepared from 10 in each tumour. RESULTSHistograms were prepared from chromosome counts in each metaphase selected ( Fig. 1-3). Each histogram shows a wide range, but there is a tendency for the majority of counts to lie around a single or a small range of numbers. These modal numbers are thought to be characteristic of the cells which form the bulk of the tumour and which are those chiefly responsible for its propagation at the time of operation. The range of chromosome counts is often greater below the modal number than above it, which may be due to loss of chromosomes in technical procedures. Table I summarises the clinical details and main chromosome findings in the present series of cases. In seminomas modal chromosome counts varied between 61 and 88. The highest number was found in case 3, where a large seminoma