Every time I attempt a transfection, my cells form into a Swiss-cheese pattern on the coverslips they are coated on. Our lab obtained new HeLa's from ATCC and they grew great and transfected fine at first, but within a few weeks they were back to clumping together, transfecting poorly, and taking on the appearance of "Swiss-cheese." Has anyone else ever had growth/transfection problems with HeLa's and if so how did you troubleshoot them?
The cells are fine and grow completely flat on the glass coverslips prior to transfection. It is only after transfection that they begin to behave strangely.
Whatever the reason, cells are getting sick. It could be mycoplasma contamination. Check if mycoplasma are contaminated. Your media, FCS, PBS etc. may be contaminated and you might get mycoplasm in your culture bottles of new cells. The other possibility is that the FCS you are using is not very good for your cells. There are some variation among batches. You might want to ask samples to get good FCS before buying it. Just in case, you do not know.
Hi Charles,
If you find that your cells are growing normally until you perform the transfection, it may be that your transfection conditions are not optimal. What type of transfection reagent are you using? Maybe you could supply some details about your transfection protocol to get a better idea about what might be happening to your cells. It's also important to seed the cells at an optimal density and use that concentration of cells for each transfection experiment. If the cell density is too low, this could also affect the health of your cells after transfection.
It is known that Hela, SiHa and Casky have low tansfection efficiency , they have the genome of HPV 18 and 16 integrated. Why don't you use HEK 293 cells?
- Badges
- Science topic