Morphological Responses of Prostatic Carcinoma to Testosterone in Organ Culture

Yang

Microscopy Res & Technique

1995

Organ culture of the human prostate began in the 1970s and was modeled after the work of Lasnitzki and her collaborators in the mouse two decades earlier. In organ culture of human prostates, one sees a rapid increase in epithelial cells and decrease in stromal cells during the first 3-5 days of culture. While modulation of many phenotypic properties occurs, these cultures provide a simple and rapid way to achieve large numbers of human prostatic epithelial cells in cultured tissues that are markedly depleted of stromal cells. There is some evidence that organ cultures are maintained in slightly better functional states in the presence of androgens; however, most of this evidence is less than quantitative. Most organ culture of prostates has been accomplished with tissues from unspecified locations within the prostate; interpretation of cultures carried out in this fashion has been less complete than would have been possible if they had been carried out from specific anatomic locations within the prostate. Careful pathological characterization of locations contiguous to the cultured tissue is mandatory if cultures are to be interpreted meaningfully.

Section: Organ Culture Of Human Prostatesmentioning

confidence: 99%

Organ culture of benign, aging, and hyperplastic human prostate

Yang

Microscopy Res & Technique

1995

“…Testosterone metabolism was thought to provide a sensitive index by which to evaluate changes in culture conditions. It has been found possible to maintain human prostatic slices in organ culture so that the morphology of the cultured expiants resembled that of the fresh tissue (McMahon et al 1972; McMahon Se Thomas 1973). To achieve greater confidence in this experimental model it is desirable to demonstrate similarities ') Present address:…”

mentioning

confidence: 99%

Testosterone Metabolism in Cultured Hyperplasia of the Human Prostate

McMahon

Butler

Thomas

1974

Acta Endocrinologica

Slices of human benign prostatic hyperplasia were maintained in organ culture in the presence of [3H]-or [14C]testosterone. Explants concentrated radioactivity from the culture medium, although this effect was depressed by the inclusion of foetal calf serum in the medium.Testosterone was metabolised to products with chromatographic mobilities corresponding to androstanediols, androsterone, dehydroepiandrosterone, and androstanedione. The principal metabolite was identified as 5\ g=a\ \ x=r eq-\ dihydrotestosterone, and small amounts of testosterone and androstenedione were also found.As culture time increased from 1 to 6 days there was a diminution in the proportion of 5\g=a\-, and an increase in 17-ketometabolites, indicating a swing to a less physiological pattern of testosterone metabolism by the aging culture. Testosterone metabolism was thought to provide a sensitive index by which to evaluate changes in culture conditions. It has been found possible to maintain human prostatic slices in organ culture so that the morphology of the cultured expiants resembled that of the fresh tissue (McMahon et al. 1972; McMahon Se Thomas 1973). To achieve greater confidence in this experimental model it is desirable to demonstrate similarities ') Present address:

“…Recent work in this laboratory has shown that carcinoma of the prostate and benign prostatic hyperplasia (McMahon, Butler and Thomas, 1972) can be cultured successfully using a modification of the grid technique (Trowell, 1959), the main innovation being that a slab of agar-gelled medium was interposed between the explant and the grid. The use of a sheet of 2% agar in 0.7 % NaCl, as an alternative to lens tissue, was originally suggested by Trowell (1959) (McMahon, 1970).…”

Section: Discussionmentioning

confidence: 99%

Oestrogen Metabolism in Cultured Human Breast Tumours

Willcox¹,

Thomas²

1972

Br J Cancer

Self Cite

Summary.-The interconversion of tritium labelled oestrone and oestradiol-17p has been investigated in human breast tumours maintained in organ culture for 3 days. Benign tumours were significantly different from scirrhous carcinomata both in the concentration of radioactivity taken up by the tissue and in the ratios of oestradiol-17p/oestrone achieved. The fact that malignant tumours were able to convert oestrone to oestradiol-17p is of interest in view of the relatively high plasma levels of oestrone in post-menopausal women.MANY of the biogenetic steroid precursors of oestradiol and testosterone are found in plasma. The transformation of precursors of low intrinsic biological activity (prehormones) into the active hormones could be a method by which certain tissues are able to adjust the hormone environment to suit their particular needs. In discussing these concepts, Baird et al. (1969) made two important points bearing on the possible role of oestrone as a prehormone: (1) In those situations where ovarian function is low or absent (as in post-menopausal and castrate women, and in men) the blood production rate of oestrone is higher than that of oestradiol-17,/; (2) In order to postulate that oestrone is a prehormone with little biological activity, the rate of conversion of oestrone to oestradiol-17,/ must be very high in certain target tissues, whereas in many other tissues the rate is low or the direction of the reaction is primarily oxidative.There is a need, therefore, to develop methods for investigating the metabolism and effects of oestrogens on tissues isolated from the complexity of the host environment. Organ culture offers one such approach. In the present study the interconversion of oestrone and oestradiol-17/1 is investigated in human breast tumours maintained in organ culture. MATERIALS AND METHODSCulture technique.-Tissue spec imens, either in the form of mastectomy specimens from the theatre or biopsy specimens from the Pathology Department, were set up in culture within 45 minutes of their removal. The specimens were washed with Eagle's basal medium (3 x 5 ml) and freed as far as possible from adhering fat and connective tissue, and necrotic areas were discarded. A razor blade was used to cut slices (4 mm2 x 0O8 mm) of the tumour tissue.Expanded stainless steel supporting grids were placed in sterile plastic petri dishes (40 mm in diameter) containing Eagle's basal medium (5 ml) supplemented with 10% foetal calf serum (Tissue Culture Services, Slough), bovine pancreatic insulin (25 ,ug/ ml), benzvlpenicillin (3 tg/ml) and streptomycin (7 ,g/ml). A block of agar-gelled medium was interposed between the supporting steel grid and the explant. This was prepared by pouring a 1.4% solution of agar in Eagle's basal medium into sterile petri dishes to a depth of 2 mm; slabs of agar were then cut to correspond to the size of the supporting grids.Culture dishes were housed in glass petri dishes (11-25 cm in diameter) which were stacked in anaerobic jars and gassed with 95% 02 and 5%/ CO2 and thereaf...