High density lipoprotein and low density lipoprotein utilization by human granulosa cells for progesterone synthesis in serum-free culture: respective contributions of free and esterified cholesterol.

Human preovulatory granulosa cells cultured in serum- and gonadotropin-free medium secreted progressively less progesterone as time elapsed. Addition of purified high density lipoproteins (HDL) as well as low density lipoproteins [very low density (VLDL) plus low density lipoproteins (LDL)] restored optimal synthesis of progesterone, and HDL was as effective as VLDL + LDL. The use of cholesterol doubly labeled lipoproteins allowed calculation of the proportions of free and esterified cholesterol converted into progesterone. Granulosa cells used either free or esterified cholesterol from VLDL + LDL. In contrast, HDL-esterified cholesterol was a poor substrate for progesterone synthesis, while HDL-free cholesterol was used preferentially. LH increased the use of both kinds of lipoproteins without changing the way in which they were used. Pretreatment of HDL by purified phospholipase A2 increased the conversion of free cholesterol into progesterone. Similar treatment of VLDL + LDL had little effect on progesterone secretion. We conclude that HDL as well as VLDL + LDL can provide cholesterol to human preovulatory granulosa cells and that utilization of HDL-cholesterol may depend on gonadotropin (LH) and enzymatic (phospholipase) regulation.