Hi! I passaged some hIPSCs yesterday, and they looked great this morning. I changed the medium (E8fn) to remove the Revitacell supplement, and they looked fine. This afternoon, I checked on them, and they had lost their morphology; they seemed to have retracted and rounded. What does this mean? Is it possible that the medium was too cold (even though I had it at RT for ~40min)? Do you know if they can recover?
Thank you!!
RT shouldn't affect this much the cell morphology; rather changes in medium pH and osmolarity (different dilution?) or use of enzymes (too much trypsin?)
Another such situation may be if you subcultured cells on dishes from another manufacturer, which have poor adhesive properties.
I agree with Felician that if the medium was kept at RT for 40min then it wouldn't be cold enough to change cell morpholgoy. It is more likely that a change in pH or osmolarity would affect the cells negatively. The point raised by Alexandr is also very true.
If none of the above options help with troubleshooting then I would suggest to double check the medium. Perhaps the E8 supplements had not been added to the E8 basal medium before feeding the cells.
Further, if you grow your cells on feeder then a different batch of feeder cells or wrong density of feeder cells can stress out the cells.
Alternatively, if you maintain iPSCs on feeder-free culture on matrix (geltrex, matrigel etc) then check those aliquots. If the concentration of matrix protein is not appropriate, it can also cause a sudden change in cell morphology.
good luck !
Hi Gemma Nomdedeu-Sancho
iPSC cell has tendency to differentiate into any other types of cell. So to minimize the iPSC differentiation you can do (I assume your iPSC medium is right)
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