Hepatocyte isolation and transplantation in the pig.
Hepatocyte transplantation (HTX) has received great expectation for the treatment of a wide spectrum of liver diseases. Considering the severe shortage of human livers for hepatocyte isolation, porcine hepatocytes are an attractive alternative to normal human hepatocytes. To develop such therapy, establishment of an efficient hepatocyte isolation and transplantation model that enables accurate assessment of safety and efficacy of HTX is extremely important. Porcine hepatocytes were isolated from a surgically removed liver segment with a four-step retrograde perfusion using dispase and collagenase. The resultant hepatocytes of > 84% viability were used for transplantation experiment in a pig model of acute liver failure induced by intravenous administration of D-galactosamine (D-gal) (0.5 mg/kg). Twenty-four hours after D-gal injection, transplantation of freshly isolated porcine hepatocytes (1 x 10(9)) was safely conducted and prolonged the survival of D-gal-treated pigs. We describe an efficient porcine hepatocyte isolation and subsequent cell transplantation in pigs with D-gal-induced liver failure.
Department of Surgery, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700-8558, Japan.
Masanobu
M
Maruyama
Toshinori
T
Totsugawa
Takemi
T
Kunieda
Teru
T
Okitsu
Norikuni
N
Shibata
Michihiko
M
Takesue
Yuzuru
Y
Kurabayashi
Mizuko
M
Oshita
Shuhei
S
Nakaji
Makoto
M
Kodama
Noriaki
N
Tanaka
Naoya
N
Kobayashi
D000818:Animals; D002469:Cell Separation / Q000379:methods; D002470:Cell Survival / Q000502:physiology; D017690:Cell Transplantation / Q000379:methods* / Q000639:trends; D017364:Collagenases; D004195:Disease Models, Animal; D010450:Endopeptidases; D005688:Galactosamine; D022781:Hepatocytes / Q000166:cytology / Q000502:physiology / Q000637:transplantation*; D008107:Liver Diseases / Q000628:therapy*; D017114:Liver Failure, Acute / Q000139:chemically induced / Q000628:therapy; D016031:Liver Transplantation / Q000379:methods* / Q000639:trends; D008297:Male; D010477:Perfusion / Q000379:methods; D015996:Survival Rate; D034421:Sus scrofa; D014183:Transplantation, Heterologous / Q000379:methods* / Q000639:trends; D016896:Treatment Outcome
D005688:Galactosamine; D010450:Endopeptidases; D017364:Collagenases; C023139:dispase