Interaction between porcine granulosa and thecal cells in steroidogenesis in an amnion dual chamber culture system

Objective: To study the interaction between granulosa and thecal cel1s in steroidogenesis in an am-nion dual chamber in comparison with the cellulose dual chamber.Method: A dual chamber culture system was designed and prepared with amnion membrane from human term placenta. The isolated porcine granulosa and thecal cells were grown on both sides of the amnion membrane, with granulosa cells in the inner chamber and thecal cells in the outer chamber. The concentrations of estradiol (E,), progesterone (P) and testosterone (T) in the culture media were mea-sured by radioimmunoassay.Results: The growth of both cells and their steroidogenic function were more active in amnion dual chamber system than in cellulose dual chamber system: (1) more T produced by thecal cells in the outer chamber, passing into inner chamber through the amnion membrane. T was used by granulosa cells as the substrate of aromatization, so that granulosa cells produced more E2 (up to 2 435 pmol/L); (2) the production of P (52. 5 μmol/L) and T (10. 2μmol/L) by granulosa cells cultured in the amnion mem-brane dual chamber system were also higher.Conclusions:The dual chamber system made of amnion mpmbrane showed better effect in studying steroidogenesis than with cellulose dual chamber system, and can be used as a model for studying paracrine regulatory interaction between granulosa and thecal cells in vitro.