摘要
目的 建立HBV感染人胎肝细胞体外培养系统。 方法 首先分离、培养人胎肝细胞,然后应用HBV阳性血清感染体外培养的人胎肝细胞;每隔2d收集上清液和肝细胞,应用ELISA、免疫组化法、原位杂交法和斑点杂交法检测上清液和细胞中HBsAg和HBV DNA。 结果 上清液中HBsAg在感染后2d～20d均可测出,以感染后4d～16d达高峰(A值在0.22左右)。免疫组化检测细胞中HBsAg呈阳性表达,原位杂交和斑点杂交检测细胞和上清液中HBV DNA也呈阳性表达。 结论 HBV在原代培养人胎肝细胞中能稳定复制和表达至少达16d。
AIM To establish a culture system of HBV-infected human fetal hepatocyte in vitro.METHODS Human fetal hepatocytes were isolated and cultured, then infected with HBV positive serum in vitro. Supernatant and cells were collected every 2 days. HBsAg and HBV DNA were detected by ELISA, immunohist-ochemistry, in situ hybridization and dot blot hybridization in cells and supernatant. RESULTS HBsAg could be detected from day 2 to day 20 after HBV-infection in supernatant and hepatocytes. Secretion of HBsAg reached submit from day 4 to 16 after HBV-infection in supernatant (A value: about 0.22). HBsAg could be detacted by immunohistochemistry in cells. HBV DNA could be detected by in situ hybridization and dot blot hybridization in cells and supernatant. CONCLUSION Primary human fetal hepatocytes are competent for infection with HBV. HBV can stably replicate and express in HBV-infected human fetal hepatocytes, and at least this replication and expression can last 16 days.
出处
《世界华人消化杂志》
CAS
2000年第4期403-405,共3页
World Chinese Journal of Digestology
基金
国家自然科学基金
No.39570166
