[Isolation of mouse primary hepatocytes by retrograde liver perfusion with catheterization via heart].

Sichuan da xue xue bao. Yi xue ban = Journal of Sichuan University. Medical science edition

PubMedID: 24527601

Bai YN, Zhang T, Wu H, Zeng Y. [Isolation of mouse primary hepatocytes by retrograde liver perfusion with catheterization via heart]. Sichuan Da Xue Xue Bao Yi Xue Ban. 2014;45(1):138-41.
OBJECTIVE
To explore the effects of retrograde liver perfusion with catheterization via heart on the isolation of primary mouse hepatocytes.

METHODS
In order to more efficiently isolate primary mouse hepatocytes, we improved the traditional two-step collagenase perfusion method. The liver perfusion catheter was inserted through right atrium and suprahepatic vena cava, and the perfusion velocity was controlled by the drip infusion with collagenase perfusate containing 10% of fetal calf serum.

RESULTS
Total success rate of catheterization in the improved group was as high as 95%, and the success rate of first at ttempt in the improved group was significantly higher than that in the traditional group (94.7% vs. 68.8%). Liver perfusion in the improved group was symmetrical with the high yield of hepatocytes up to 1.07 x 10(6) per gram of mouse weight and 92.16% of average cell vitality, which were higher than those in the traditional group.

CONCLUSION
The retrograde liver perfusion through the heart is a simple and easy-to-learn method to isolate mouse primary hepatocytes, which also could guarantee the satisfactory yield and vitality of primary hepatocytes.