[Primary culture of human trophoblastic cells and their susceptibility to human cytomegalovirus].

ABSTRACT

OBJECTIVE: To determine the susceptibility of primary human trophoblastic cells (HTCs) to human cytomegalovirus (HCMV) infection. METHODS: 30 villus of healthy pregnant women were digested with a mixture of 0.25% trypsin and 0.2% DNase. The digestion was then filtered by a 200 mesh stainless steel sieve. HTCs were purified by Percoll gradient centrifugation and repeat purification with trypsin during cell passage. HTCs were identified by cell morphology and immunofluorescence. PCR, Western blot and immunofluorescent staining were performed to detect HCMV gene and protein 72 hours post infection of HCMV. RESULTS: The combination of 0.25% trypsin and 0.2% DNase with Percoll gradient centrifugation isolated and purified primary HTCs. More than 90 percent cells expressed CK-7 related antigen. The results of PCR, western blot and immunofluorescence showed positive expressions of CMV gB and virus proteins. CONCLUSION: HTCs is susceptible to HCMV infection, which suggests a possible route of HCMV infection at the fetal-maternal interface.