A carrier-free cell superfusion system for the study of androgen formation from precursor steroids by isolated rat testis Leydig cells under steady-state conditions

A cell superfusion system is presented in which cells can be kept in suspension if there is an equilibrium between cell sedimentation velocity and superfusion medium flow velocity. Since the cross-section area of the central core increases gradually, theoretical considerations and experimental results demonstrate that single isolated cells as well as small cell aggregates are not able to leave the superfusion chamber. The whole apparatus is constructed from glass to avoid adsorption of steroids. As an application of the system, androgen formation from progestin precursors by isolated rat testis Leydig cells is shown. Under steady-state conditions, constant concentrations of substrates, intermediates, and products are measured in the collected effluent. In contrast to conventional cell incubation, constant testosterone formation rates and linearly increasing cumulative testosterone production are achieved.