I'm working with mamma cells, some of which grow slowly and others grow fast. I work mainly on the S phase, so I have to prepare my cells one day (24 hours) before the experiment and then run the experiment. I have doubt that all cells can enter S phase after 24 hours.
Dear Hamed, your doubt are correct, not all the cells once plated will enter S phase at the same time. One way to synchronize the cells is to do a serum-starvation. That is, once you plated the cells, let them attach (16-24 h depending on cells) then you put in medium without serum for 4-6h (again, depending on cell) and they will go in G0 phase, synchronizing themselves. You add then fresh medium with serum and after 16-24h they will be all in S phase. I use this strategy to increase transfection efficiency...
Dear Hamed, your doubt are correct, not all the cells once plated will enter S phase at the same time. One way to synchronize the cells is to do a serum-starvation. That is, once you plated the cells, let them attach (16-24 h depending on cells) then you put in medium without serum for 4-6h (again, depending on cell) and they will go in G0 phase, synchronizing themselves. You add then fresh medium with serum and after 16-24h they will be all in S phase. I use this strategy to increase transfection efficiency...
I think u can carry cell cycle analysis using PI and flow cytometry on different time (6, 12 and 24hr), and see the percentage of cell in the S phase in every time, it may help.
- Badges
- Science topic