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目的 研究TGF β(transforminggrowthfactorbeta)对体外分离纯化培养的滋养细胞增殖及功能的影响。方法 将Percoll梯度离心法分离纯化的滋养细胞接种于培养板 ,2 4h贴壁后换液。加药组分别加含 2 5、5 0、10 0 μg/LTGF β的 199培养液 ,对照组培养液中无TGF β ,继续培养2 4h ,在光镜下观察各组细胞生长情况 ,用MTT法测定加药和对照组OD值 ;用酶联法测定培养液中人绒毛膜促性腺激素 (hCG)的浓度 ,并取培养板中的盖片进行hCG免疫组化染色及图像分析。结果 加药组细胞生长密度及MTT法测得的OD值小于对照组 ;培养液中hCG浓度及盖片免疫组化的OD值均明显小于对照组。结论 TGF β可抑制体外培养的滋养细胞生长及分泌hCG的功能。
Objective To investigate the effects of transforming growth factor βbeta (TGF β) on the proliferation and function of trophoblasts isolated and purified in vitro. Methods The trophoblast cells isolated and purified by Percoll gradient centrifugation were inoculated into the culture plate and were changed after adhering to the plate for 24 h. In the drug-treated group, 199 medium containing 25, 50 and 100 μg / L of TGFβ was added respectively. In the control group, no TGF-β was added in the culture medium for 24 hours. The cell growth was observed under light microscope, Method was used to determine the OD value of dosing group and control group. The concentration of human chorionic gonadotropin (hCG) in the culture medium was determined by enzyme-linked immunosorbent assay (ELISA). The hCG immunohistochemical staining and image analysis were performed on the cover plate. Results OD value of cell growth density and MTT assay in dosing group was smaller than that in control group. The concentration of hCG in culture solution and the OD value of immunohistochemistry were significantly lower than those of control group. Conclusion TGF β can inhibit the growth of trophoblasts and the secretion of hCG in vitro.