hello, this is soyoung hwang.
Recently, i tried to generate organoids from iPSC with Aggrewell
However, on D4~D6, it starts to die. The lining around the organoid surface became rough, and lots of single cells were found in the media.
Actually, on the very first day of aggregation, several cells were found around organoids in the aggrewell.
I wonder it might be the problem of cell condition.
any idea ?
Kindly greeting, Soyoung.
Hi,
The situation observed in the photos is typical during using an aggrewell plates for embryoid bodies forming. Around the cells aggregates, there are dead cells that did not form embryoid bodies and dead cells that detached from the embryoid bodies. iIPS cells are cells that grow and like growing in colonies. For use of aggrewell plate, the colonies are broken down into single cells. Dissociation of iPS colonies into single cells is stressful for iPSCs. After the transfer of the embryoid bodies from aggrewell plate to the culture plate, dead cells will be removed during the media exchange.
Good luck,
Justyna Augustyniak
Cell quality is important, in our experience, factors such as The number of passages, the quality of the single cell suspension, the presence of diferentiated cells and the level of confluency at culture might have relevant impact in the problems ypu are facing. Good luck!
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