Progesterone-dependent secretion of macrophage colony-stimulating factor by human endometrial stromal cells of nonpregnant uterus in culture.

Abstract

Recent studies have suggested that macrophages colony-stimulating factor (M-CSF), a hematopoietic glycoprotein essential to the proliferation and differentiation of mononuclear phagocytes and their progenitor cells, is also involved in the reproductive process in mice and humans. In this study, we examined, by enzyme-linked immunosorbent assay, the supernatants of stromal cell-enriched fraction (SF) of human nonpregnant endometrium for the presence of M-CSF during culture with progesterone (P) or estrogen. The bioactivity of M-CSF was assessed in a colony-forming assay of murine bone marrow cells. In addition, the M-CSF level in the culture supernatant of SF further purified by subculture, of epithelial cell-enriched fraction purified from human endometrium, and of peripheral blood lymphocytes, including about 10% monocytes, was examined with or without P, because SF is contaminated by epithelial cells and macrophages, both of which are suggested to secrete M-CSF. During 2-week culture, the level of M-CSF in the supernatants of SF cultured with P was markedly higher than that of control culture and estrogen-treated culture on any day tested, except for the first 2 days. P had a dose-dependent effect on M-CSF production by SF. Estrogen also enhanced M-CSF production by SF, but did not show dose dependency. The SF culture supernatants showed a colony-forming activity that was completely blocked by neutralizing anti-M-CSF antibody. SF subcultured three times, which was confirmed to be of more than 99% purity, secreted M-CSF in a P-dependent manner. M-CSF was also detected in the culture supernatants of epithelial cell-enriched fraction and peripheral blood lymphocytes, but P-dependent M-CSF production was not shown in these cultures. These results suggest that human endometrial stromal cells themselves can secrete bioactive M-CSF in a P-dependent manner in vitro, indicating that the M-CSF reported to be present in human endometrium is secreted in part by stromal cells and may play a role in the regulation of endometrial function under P control.

The Role of Cytokines in Human Endometrium: The Inhibitory Effect of IL-1 and TNF α on <I>in Vitro</I> Decidualization and mRNA Expression of M-CSF, SCF and LIF in the Human Endometrium

“…It has become apparent that endometrial cells themselves can produce some cytokines.Kauma et alreported that the human endometrium and decidua expressed M-CSF mRNA,and the M-CSF protein was found in extracts from the endometrium [27].By means of ELISA with specific anti-M-CSF antibodies,we have reported that human endometrial stromal cells produce M-CSF and the production was dependent on progesterone but not on estrogen added to the cultures [28].In the present study,with Northern blot analysis,we confirmed a 4-kb mRNA for M-CSF in human endometrial stromal cells,and revealed that M-CSF mRNA expression was increased with progesterone supplementation (Fig.4).In these culture experiments,the level of mRNA expression in the cells was dependent on the amount of progesterone added to the cultures (Fig.5).…”

Concentration of Macrophage Colony-Stimulating Factor (M-CSF) in Bovine Peripheral Blood during Pregnancy

“…In human nonpregnant endometrial tissues, the tissue-extractable level of immunoreactive M-CSF in secretory endometrial tissue is higher than that in proliferative endometrial tissue [15]. Furthermore, progesterone has a dose-dependent effect on M-CSF production by the stromal cell-enriched fraction of the human non-pregnant endometrium in vitro [13]. M-CSF has several isoforms, and bovine M-CSF is expressed mainly as 3-and 4-kb transcripts; the 3-kb mRNA encodes the membrane-bound form, and the 4-kb mRNA encodes the secreted form.…”

“…A number of researchers have reported that M-CSF and its receptor are expressed in the female reproductive tract and that M-CSF plays an important role in pregnancy in mice [1,2,[21][22][23], humans [8,[11][12][13], pigs [35] and cows [3,17,19]. Furthermore, M-CSF concentrations in maternal peripheral blood during the estrous cycle and pregnancy have been reported in humans and mice.…”

“…In contrast, human endometrial, decidual and placental tissues have been found to express M-CSF mRNA and have demonstrated extractable immunoreactive M-CSF [8,12,13]. Human M-CSF, in a concentration-dependent manner, increases the secretion of human chorionic gonadotropin from primary cultured human cytotrophoblastic cells and a human placental cell line [27].…”

Danazol regulates the functions of normal human endometrial stromal cell subpopulations by modifying endometrial cytokine networks

“…The inhibitory effects of danazol on ESC activation into G-CSF-secreting cells are opposite to the effects of M-CSF, which induces ESC activation into G-CSF-secreting cells. M-CSF is produced in the human endometrium, particularly by decidual cells (33,34). IL-8 secretion by ESCs is regulated by endometrial IL-1 and IL-1 receptor antagonist (31).…”