Neonatal Rat Cardiac Myocyte Isolation

[shyn]

Dear all,

I've recently started isolating cardiac myocytes from 1-2day old Wistar pups by the trypsin digestion method with about 10-15 cycles of 6-8min digestion each time.

However, I've been unsuccesful in getting a high cell yield. The amount isolated from 15-20 pups each time is only sufficient to plate in 1 T75 flask.
Moreover, in the first few digestion cycles (cycle 1 -cycle 5); i'm barely able to collect any cells; and there's this plasma/gel-like substance that appears with the minced tissue. Wondering if this is normal?

Would gladly appreciate any tips/advice that you guys might be able to offer!

Thanks!

shyn