WI38 cells - problems with transfectations of this ce (Jul/06/2004 )
I have some problem with lipofectamine transient transfection using WI38 line, Does anybody conduct transfection of WI38 ?
To find such information, the best source is Medline. Here are two examples:
WI38, grown to 60-95% confluency, were detached using trypsin + EDTA (<1 min, room temperature). The cells were washed with trypsin neutralizing solution (Cascade) and pelleted. To avoid electroporating in the presence of heparin (present in supplemented Medium 200), the cells were suspended with RPMI1640 + 10% NuSerum IV to a concentration of 0.8-1.0X107 cells/ml and kept on ice. Samples of cell suspension (100-400ul) were mixed with plasmid DNA (in 5 ul water per 100 ul cell suspension), warmed briely to room temperature, and electroporated in a 4-mm cuvette (BioRad, Hercules, California) at 250 V. Capacitance was set to 250 uF per 100 ul cell suspension, giving a time constant of about 30 msec.
Briefly, 10^7 cells were mixed with 50 ug of DNA and were electroporated at 260 V, 960 microfarads (Gene PulserTM, Bio-Rad). Transfected cells were split 48 h later and were maintained in complete medium containing 400 ug/ml G418.