[Microbiology] I don't know why!!!

[tuffwave]

I'm working about lactic acid bacteria.
I used 20% glycerol+20% glucose solution for preventing the damage by
freezing.
In lab-scale, this process is normal.
But when this process is scale-up (250L main fermentation->suspension
using 15 L anti-freezing reagent), lactic acid bacteria was dead over
90%.
I don't know why cell death by scale-up was happened.

Help me!!!