Kariya M, Kanzaki H, Hanamura T, Imai K, Narukawa S, Inoue T, Hatayama H, Mori T
Department of Gynecology and Obstetrics, Faculty of Medicine, Kyoto University, Japan.
J Clin Endocrinol Metab. 1994 Jul;79(1):86-90. doi: 10.1210/jcem.79.1.8027260.
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.
近期研究表明，巨噬细胞集落刺激因子（M-CSF）是一种对单核吞噬细胞及其祖细胞的增殖和分化至关重要的造血糖蛋白，它也参与小鼠和人类的生殖过程。在本研究中，我们通过酶联免疫吸附测定法，检测了人非孕子宫内膜富含基质细胞的组分（SF）在与孕酮（P）或雌激素共同培养期间上清液中M-CSF的存在情况。通过小鼠骨髓细胞集落形成试验评估了M-CSF的生物活性。此外，由于SF被上皮细胞和巨噬细胞污染，而这两种细胞均被认为可分泌M-CSF，因此我们检测了经传代进一步纯化的SF、从人子宫内膜纯化的富含上皮细胞的组分以及外周血淋巴细胞（包括约10%的单核细胞）在有无P存在情况下培养上清液中的M-CSF水平。在为期2周的培养期间，除了最初2天外，在任何测试日，与P共同培养的SF上清液中M-CSF的水平均显著高于对照培养和雌激素处理培养的上清液。P对SF产生M-CSF具有剂量依赖性作用。雌激素也可增强SF产生M-CSF，但未显示出剂量依赖性。SF培养上清液表现出集落形成活性，该活性被中和性抗M-CSF抗体完全阻断。传代三次且纯度超过99%的SF以P依赖性方式分泌M-CSF。在富含上皮细胞的组分和外周血淋巴细胞的培养上清液中也检测到了M-CSF，但在这些培养物中未显示出P依赖性M-CSF产生。这些结果表明，人子宫内膜基质细胞自身可在体外以P依赖性方式分泌生物活性M-CSF，这表明据报道存在于人子宫内膜中的M-CSF部分由基质细胞分泌，并且可能在P调控下的子宫内膜功能调节中发挥作用。
