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目的 建立HBV阳性血清体外感染HepG2 细胞的实验方法。方法 培养HepG2 细胞传至6孔板中,2 4h后进行HBV阳性血清体外感染HepG2 细胞的实验。感染组用HBV阳性血清,阴性对照组用HBV阴性血清,空白对照组用DMEM培养基。实验开始后HepG2 细胞继续孵育2 4h ,而后用0 0 1mol LPBS清洗8次后加入2 %DMEM培养液。收集PBS第8次洗液,收集PBS洗后每隔12h各孔细胞培养上清。ELISA检测细胞培养上清中的HBsAg。PCR检测细胞培养上清和HepG2 细胞中的HBVDNA。结果 感染组在PBS洗后12h的细胞培养上清中ELISA检测HBsAg呈阳性。PCR检测显示感染组细胞培养上清和HepG2 细胞中HBVDNA呈阳性,阴性对照组和空白对照组HBVDNA呈阴性。结论 HBV阳性血清进行HBV感染体外培养HepG2 细胞是可行的。
Objective To establish an experimental method for the in vitro infection of HepG2 cells with HBV positive sera. Methods HepG2 cells were cultured and passaged into 6-well plates. After 24 h, HepG2 cells were infected with HBV-positive serum in vitro. HBV-positive sera were used in the infected group, HBV-negative sera in the negative control group and DMEM medium in the blank control group. After the start of the experiment, HepG2 cells were incubated for 24 hours, then washed with 0 0 1mol LPBS for 8 times and then added with 2% DMEM. Collect the 8th washings of PBS and collect the cell culture supernatants from each well every 12h after washing with PBS. ELISA to detect HBsAg in cell culture supernatant. PCR detection of HBVDNA in cell culture supernatant and HepG2 cells. Results In the infection group, positive HBsAg was detected by ELISA in the cell culture supernatant of 12h after washing in PBS. PCR detection showed that HBVDNA was positive in the cell culture supernatant and HepG2 cells in the infected group, but negative in the negative control group and the blank control group. Conclusion Hepatitis B virus (HBV) positive serum is feasible to culture HepG2 cells in vitro.