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Table 1 Frequent causes unsuccessful hepatocyte isolation.

From: Improved isolation of murine hepatocytes for in vitro malaria liver stage studies

Step Problem Possible Reason Solution
Perfusion Poor cell dissociation Poor digestion Make sure the Digestion solution is at 37°C
   Tissue disruption Lower down perfusion speed
  Cell death Excessive collagenase digestion Reduce the time/volume of perfusion with Hepatocyte Liver Digest Medium
Cell isolation Low recovery To many cells loaded on the gradient If using more than three lobes, increase the number of gradient tubes
   Gradient disturbance Setup new gradient, making sure the layers do not mix
   Altered gradient densities Check that the gradient centrifugation temperature is kept at 20°C and Percoll solutions at room temperature