Effects of blood monocytes and lymphocytes on progesterone secretion by granulosa cells in the pig

Abstract

We studied the effects of autologous and nonautologous co-cultures of porcine blood monocytes and lymphocytes with granulosa cells on progesterone secretion. Eight prepubertal crossbred gilts were ovariectomized, and the granulosa cells were collected, plated at 2.5 × 10 5 cells/ml and allowed to attach. Blood was obtained from the same eight gilts, and the mononuclear cells were separated by density gradient centrifugation. Monocytes were separated from lymphocytes by adherence to plastic. Adherent monocytes, lymphocytes and a 1:1 mixture of monocytes + lymphocytes were added to granulosa cell cultures and incubated for 48 h. Progesterone secretion into the media was measured. In addition, blood cell alloreactivity was studied in these co-cultures by measuring uptake of 3H-thymidine. The co-culture of adherent monocytes or monocytes + lymphocytes with granulosa cells increased ( P <.05) progesterone secretion as compared with granulosa cells cultured alone. However, co-culture of lymphocytes with granulosa cells did not have a significant effect. No difference was observed between autologous and nonautologous cell cultures in blood cell proliferation or granulosa cell progesterone secretion. In conclusion, blood monocytes influence progesterone secretion by granulosa cells. In addition, there was no difference in the ability of autologous and nonautologous blood cells to stimulate progesterone secretion by granulosa cells. No alloreactivity was observed using nonautologous immune cells with granulosa cells.