A study on high efficiency in vitro amplification of adenoviral expression vector

Objective To explore an effective way for increasing amplification efficiency of adenoviral expression vector in vitro. Methods Approximately 70%-80% of confluent human embryonic kidney cells (HEK293) were co-cultured with recombinant adenoviral vector (Ad5CMVLacZ) for 36-48 h. X-gal histochemistry was performed to identify the transfection and expression of Ad5CMVLacZ. Results Under the normal condition, 70%-80% of confluent HEK 293 cells were logarithmically grown. Afler 36-48 h of co-culture with Ad5CMVLacZ, the shape of HEK 293 cells was gradually changed into spherical, and congregated into grape-like aggregation which is the typical cytopathic effect of Ad virus transfection of host cells. At this stage aboat 90% of transfected HEK 293 cells were staimed blue color by X-gal histochemistry. Conclusion The method is an effective way for amplification of adenoviral expression vector in vitro with paying more attention to ascertaining the time of transfectisn of HEK 293 cells with the viral vector and the time for harversting the transfected cells so as to obtain enough viral vector for gene therapy.