I have several criovials with adipose steam cells. I thawed the frozen cells and cultured them with DMEM and 5% platelet lysate on Corning T75 flasks, but they won't attach nor grow on the flask. I tried increasing the platelet lysate to 15% but didn't work apparently. I'm considering changing to FBS but it would interfere with the purpuose of the research. What else could I try? How long does it take for this cells to attach?
PD: We don't use heparin, we let the lysate defrost, then centrifugate at low speed to separate the fibrin clot and then use the liquid remaining lysate.
Also today I performed a cell viability assay with trypan blue and apparently ther is a good percentage of viable cells.
I would really appreciate any assistance, thank you.
Hi Juan,
First you should coat T-75 with Fibronectin (1mg/ml) and let it dry for 45-60 minutes.
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