Development of highly functional long-term culture method of liver slice embedded in agarose gel for bioartificial liver.
It is difficult to a produce highly functional bioartificial liver (BAL) using only hepatocytes, because it is believed that liver-specific three-dimensional structure is necessary to maintain high function for BAL. But it is difficult to construct a culture system with liver-specific three-dimensional structure in vitro. To realize a highly functional culture system with liver-specific three-dimensional structure, we developed a culture system using liver slices that keep liver-specific architecture, such as liver lobule and hepatic microvascular system. Liver slices were embedded in agarose gel to maintain them under a moist and three-dimensional environment. We examined the viability and function of liver slices by using various shapes of agarose gel. Liver slices were cultured 1) under stationary condition (control), 2) directly embedded in gel, and 3) embedded in cylindrical gel for good drainage of medium and ventilation of air. The viability and function of the incubated liver slices were evaluated by LDH leakage, histomorphology, and immunohistochemistry. At 10 days, the morphological condition and function of liver slices embedded in cylindrical gel were maintained better than liver slices directly embedded in gel or in the stationary condition. We suggest that high functionality and morphological condition of liver slices could be maintained by embedding in cylindrical gel. In the future, it is possible that this method could be used to develop a highly functional bioartificial liver.
Nonaka
Hideki
H
Department of Functional Polymer Science, Faculty of Textile Science and Technology, Shinshu University 3-15-1 Tokida, Ueda 386-8567, Japan.
Ise
Hirohiko
H
Sugihara
Nobuhiro
N
Hirose
Shinichi
S
Negishi
Naoki
N
Kondo
Yoshiyuki
Y
Akaike
Toshihiro
T
D000818:Animals; D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; D002470:Cell Survival; D046508:Culture Techniques / Q000379:methods*; D004268:DNA-Binding Proteins; D005164:Factor IX / Q000378:metabolism; D051557:Hepatocyte Nuclear Factor 4; D007150:Immunohistochemistry; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008099:Liver / Q000378:metabolism / Q000502:physiology*; D019164:Liver, Artificial; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D008833:Microcirculation; D010750:Phosphoproteins / Q000378:metabolism; D012685:Sepharose / Q000737:chemistry*; D013997:Time Factors; D014157:Transcription Factors / Q000378:metabolism
D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; D004268:DNA-Binding Proteins; D051557:Hepatocyte Nuclear Factor 4; D010750:Phosphoproteins; C501970:Tcfl4 protein, mouse; D014157:Transcription Factors; D005164:Factor IX; D012685:Sepharose; D007770:L-Lactate Dehydrogenase