人滋养层细胞内丙型肝炎病毒样颗粒的免疫电镜形态学研究

Immunoelectron microscopy observation of HCV-like particles in the human cytotrophoblasts cultured in vitro

目的:应用免疫电镜技术观察体外培养的人胎盘滋养层细胞感染丙型肝炎病毒(HCV)情况。方法:采用胰蛋白酶消化法及Percoll密度梯度分离法分离培养人胎盘组织中滋养层细胞,以HCVRNA阳性血清对滋养层细胞进行体外感染实验,胶体金免疫电镜检测滋养层细胞内的HCV病毒颗粒,逆转录聚合酶链反应(RT-PCR)定性检测滋养层细胞内HCVRNA。结果:体外培养的人胎盘滋养层细胞内可观察到胶体金颗粒吸附的病毒样颗粒,形态及直径符合HCV特点。RT-PCR定性检测结果阳性,进一步证实所观察到的病毒样颗粒为HCV。结论:HCV可感染体外培养的人胎盘滋养层细胞。

Objective ： To provide the morphological evidence of HCV infection of the human cytotropnoblasts cultured in vitro by irnmunoelectron microscopy . Methods： Human placentae were digested with trypsin and centrifuged with percoll density gradient to obtain cytotrophoblasts. The cytotrophoblasts were incubated in DMEM-HG medium with HCV-RNA-positive serum. Prepare specimen for immunoelectron microscopy and observe under transmission electron microscope （TEM）. Qualitative RT-PCR was used to convince the results. Results： Some virus-like particles conjoined with golden granules, 45 -65 nm in diameter, with clear envelope, could be observed, which were coincident with the characters of HCV particle. Positive result of qualitative RT-PCR confirmed the observation. Conclusion： Cytotrophoblasts cultured in vitro could be infected by HCV.