Hello fellows,

I use H1299 cells for my experiments for 3 years, while there are someting wrong happened to my cells and frustrating me so hard recently.

The following figure 1 and 2 are the photos I took after 2 days of cell thawing from the liquid nitrogen, the cell morphology seems good and healthy,

and the figure 3 and 4 are the photos I took after 6 to 7 passages, the morphology is weird, like the cells are shrinking and very unhealthy. The cell debris can also be observed under microscopy field.

The bacterical and mycoplasma contamination was excluded and the incubator was autoclaved and calibrated recently.

I also maintain another cell line (SAS, an oral cancer cell) but it seems nothing strange happened.

My collegues use same batch of culture materials (FBS, trypsin, p/s antibiotic, base medium...) with me

This problem has never happened before and bothered me for over three months!

Any suggestions or clues are welcome and appreciated