We have prefered 15 minutes, but we want to try shorter times. Do you think that shorter time intervals than 15 minutes can be better for isolation? Any suggestions for the optimal incubation time?
This depends. Are you trying to use the collagenase to free up cells before passage?
To expand on the previous answer, it depends on the intent. Are you trying to use collagenase to free up cells from tissue culture plates or from collagen gels, or are you trying to isolate HUVEC from umbilical cords?
We usually incubate collagenase for 12 minute, longer time may result more heterogene cell assambly (eg.:muscle cells as well) in the cell culture.
For maximum detachment 10 minutes incubation in 0.2% collagenase should be done.Longer incubation could result in the detachment of SMC(smooth muscle like cells) which could take over your HUVEC culture.
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