| Abstract: | A procedure is described for separating guinea-pig prostatic tissue into viable epithelial and stromal fractions. Epithelial cells were isolated using 0.1% protease, but this method resulted in significant damage to the stromal cells. However, using a mechanical tissue fractionation technique, a viable stromal matrix consisting predominantly of confluent sheets of smooth muscle cells and intervening collagen fibres was obtained. Although this method selectively spilled-out the epithelial cells, the majority were non-viable and therefore not suitable for receptor studies. Electron microscopy confirmed that cell architecture and organelle morphology were well preserved in both the enzymatic epithelial preparation and the mechanically prepared stroma. Saturation analysis studies indicated that the concentration of high affinity (Kd approximately 0.15 nM) oestrogen receptors was approx. 10-times greater in the separated stroma than in the epithelial fraction. In contrast, the concentration of androgen receptors (Kd approximately 2.2nM) was almost 2-fold greater in the epithelial than in the stromal fraction. These findings suggest that oestrogen, either independently or in association with androgen, may play a role in the growth and development of the stromal component of the guinea-pig prostate. |
