Can anyone suggest a good method / composition of enzymes that allow for an efficient singularization / separation of cells from a mesenchymal stem cell aggregate / spheroid?
I tried to detach cells from one another in order to receive a single cell suspensions after 3D culitvation in an aggregate / spheroid with: collagenase (tried severalconcentrations), Accutase, Accumax, Trypsin, TrypLE, Elastase, Dispase and mixtures of these. None of it worked very well. Recovery is max. 50 %.
I am working with primary MSCs only and they produce a massive amount of ECM or at least cell cell connections. Its crazy... I know from other kinds of stem cells or stem cell cell lines that usually enzymes like Accumax / Accutase / Trypsin or collegenase will do the trick. But I did not have success with them on MSCs.
Any suggestions?
Thanks a lot.
Look at accutase or accumax but only works for 100um spheroids, not above
Dominik Egger hello,
finally you found a good strategy for dissociate spheroids?
I'm working with primary human MSCs and i have tried to detach the cells with trypsin, tryple select and accutase, but the viability post treatment its still very low.
If you have any advice i would really appreciate it...
Thanks!!
Paloma Fuentes Robert I didnt find a "perfect" method, but wtih collagenase IA + Hyaluronidase we receive about 60-70% of cells, with collagenase IA+Elastase or collagenase II we receive around 50-60%. In these conditions we observe >90% viability after dissociation. With other enzymes(accumax,Accutase etc) we receive
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