Table 1 |
|||
|
Frequent causes unsuccessful hepatocyte isolation. |
|||
| 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 |
||
|
|
|||
|
Gonçalves et al. Malaria Journal 2007 6:169 doi:10.1186/1475-2875-6-169 |
|||
