[Effects of macrophage colony-stimulating factor on production of estradiol and progesterone by human luteinized granulosa cells in vitro and detection of macrophage colony-stimulating factor receptor on human luteinized granulosa cells].
OBJECTIVE
To investigate the effects of macrophage colony-stimulating factor (M-CSF) on estradiol and progesterone production by human luteinized granulosa cells in vitro and to detect M-CSF receptor in human granulosa cells.


METHODS
Human luteinized granulosa cells (LGC) were isolated from follicular fluid of superovulated infertile patients undergoing intracytoplasmatic sperm injection. Some of the LGC were used for detecting M-CSF receptor by immunocytochemical staining (ABC method). Most of them were cultured with HAM's F-10 medium plus various concentration of M-CSF (0, 10, 25, 50, 100, 250 ng/ml) in the presence or absence of follicle stimulating hormone (FSH). Media were collected at 72 hours after culture and estradiol (E(2)) and progesterone (P) in media were measured by enzyme immunoassays.


RESULTS
About 80% of the LGC presented a positive M-CSF receptor staining with the immunochemical signal on cell membrane. After 72 hours culture of LGC, in absence of FSH, the baseline concentration of E(2) and P were (2,185 +/- 189) pmol/L, (3,157 +/- 401) nmol/L respectively; with the increasing dose of M-CSF from 10 to 100 ng/ml, the concentrations of E(2) increased from (2,789 +/- 365)pmol/L to (4,282 +/- 318) pmol/L, and those of P increased from (4,256 +/- 595) nmol/L to (7,789 +/- 828) nmol/L. In presence of FSH (75 IU/ml) the baseline concentrations of E(2) and P were (5,045 +/- 486) pmol/L and (8,667 +/- 923) nmol/L respectively; while the 10 - 100 ng/ml M-CSF was added the concentrations of E(2) and P were from (6,567 +/- 673) to (8,373 +/- 935) pmol/L and from (10,999 +/- 985) to (14,990 +/- 1,158) nmol/L respectively. Thus M-CSF caused a significant dose dependent increase of estradiol and progesterone production (P < 0.05), and M-CSF+ FSH further stimulated higher production of E(2) and P by LGC than M-CSF or FSH alone (P < 0.05).


CONCLUSIONS
M-CSF receptor was expressed on membrane of human LGC. M-CSF enhances estradiol and progesterone production by LGC and FSH has an additional or synergetic effect with M-CSF.
Department of Gynecology, Obstetrics and Gynecology Hospital, Zhejiang University School of Medicine, Hangzhou 310006, China.
Zhifen
Z
Zhang
Ali
A
Salmassi
Liselotte
L
Mettler
D000328:Adult; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D004958:Estradiol / Q000096:biosynthesis*; D005260:Female; D005640:Follicle Stimulating Hormone / Q000008:administration & dosage / Q000494:pharmacology*; D006107:Granulosa Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D016173:Macrophage Colony-Stimulating Factor / Q000008:administration & dosage / Q000494:pharmacology*; D011374:Progesterone / Q000096:biosynthesis*; D016186:Receptor, Macrophage Colony-Stimulating Factor / Q000032:analysis*
D011374:Progesterone; D004958:Estradiol; D016173:Macrophage Colony-Stimulating Factor; D005640:Follicle Stimulating Hormone; D016186:Receptor, Macrophage Colony-Stimulating Factor