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


CONCLUSIONS
HTCs is susceptible to HCMV infection, which suggests a possible route of HCMV infection at the fetal-maternal interface.
Department of Obstetric & Gynecologic, West China Second Hospital, Sichuan University, Chengdu 610041, China.
Shu
S
Zhou
Li-qun
LQ
Dong
Lin-bo
LB
Guan
Ping
P
Yu
D003587:Cytomegalovirus / Q000254:growth & development / Q000502:physiology*; D003586:Cytomegalovirus Infections / Q000821:virology; D004198:Disease Susceptibility; D005260:Female; D006801:Humans; D011247:Pregnancy; D061251:Primary Cell Culture; D014327:Trophoblasts / Q000166:cytology* / Q000821:virology*