Isolation and cultivation of porcine hepatocytes for extracorporeal artificial liver support system.
OBJECTIVE
To develop procedures for the successful harvesting of large quantities of viable and functional pig liver cells from abattoir organs.


METHODS
The procedure included partial liver lobe retrograde perfusion and mechanical/enzymatic digestion of the liver tissue, followed by separation of the hepatocytes, based on size and density, from contaminating cell types.


RESULTS
Digestion of the partial liver lobe resulted in an average yield of 1.39 x 10(9) cells (9.9 x 10(8) cells/g liver) with an average viability of 92.5%. The yield and viability of cells were improved by dispase/collagenase resultant digestion. The emergence of blebby cells was blocked by supplying oxygen to the cell isolation buffers. Isolated hepatocytes seeded onto polystyrene surfaces remained viable and functional at a level comparable to that of rat hepatocytes, although their function decreased over time.


CONCLUSIONS
Adult pig hepatocytes can be harvested with high yields and retain viability and differentiated function using this method. Abattoir pig livers can be an excellent source of hepatocytes for use as the biological component of artificial liver assist devices.
Zhou
X
X
Bionic Design Group, National Institute for Advanced Interdisciplinary Research, 1-1-4 Higashi, Tsukuba, Ibaraki 305-8562, Japan.
Liu
L
L
Kano
J
J
Mukaiyama
T
T
Tokiwa
T
T
D000418:Albumins / Q000378:metabolism; D000641:Ammonia / Q000378:metabolism; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002470:Cell Survival; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D019164:Liver, Artificial; D051381:Rats; D013552:Swine; D013997:Time Factors
D000418:Albumins; D000641:Ammonia