I am generating iPSCs from peripheral blood using Sendai Cytotune 2.0 kit. The cells attached to the surface when shifted to Matrigel coating and formed nice aggregates in the transduced wells by Day 6. However since then, they don't seem to be growing or forming colonies as they should by Day 12-13. They are neither dead or proliferating. I tried adding ROCK inhibitor for 24 hours too, with no luck.
Any troubleshooting ideas? When shifting to Matrigel, I had not spun down the cells and resuspended in Stempro without cytokines as recommended in their protocol. I had shifted the entire cells containing cytokine media onto Matrigel, then slowly removed the cytokines over 2 days, with half media changes. Could that affect this?
Gabriel Liguori thanks! Although the aforementioned problem solved itself, ie. a few colonies did attach and they eventually grew into more colonies. But thanks a lot for the info, I did not know about tissue-specific hydrogels.
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