Rat hepatocyte culture model of macrosteatosis: Effect of macrosteatosis induction and reversal on viability and liver-specific function

JOURNAL OF HEPATOLOGY

卷:59

Article

Nativ, Nir I.¹  Yarmush, Gabriel¹  Chen, Alvin¹  Dong, David¹  Henry, Scot D.²  Guarrera, James V.²  Klein, Kenneth M.³  Maguire, Tim¹  Schloss, Rene¹  Berthiaume, Francois¹  Yarmush, Martin L.^1,4 

[1] Rutgers State Univ, Dept Biomed Engn, Piscataway, NJ 08854 USA

[2] Columbia Univ Med Ctr, Ctr Liver Dis & Transplantat, Dept Surg, New York, NY USA

[3] UMDNJ New Jersey Med Sch, Dept Pathol & Lab Med, Newark, NJ USA

[4] Massachusetts Gen Hosp, Ctr Engn Med Surg Serv, Boston, MA 02114 USA

关键词: Steatosis;    Triglyceride;    Albumin;    Urea;    Bile;    Liver transplantation;    Lipid metabolism;   

DOI  :  10.1016/j.jhep.2013.07.019

来源: Elsevier

【 摘 要 】

Background & Aims: A common cause of liver donor ineligibility is macrosteatosis. Recovery of such livers could enhance donor availability. Living donor studies have shown diet-induced reduction of macrosteatosis enables transplantation. However, cadaveric liver macrosteatotic reduction must be performed ex vivo within hours. Towards this goal, we investigated the effect of accelerated macrosteatosis reduction on hepatocyte viability and function using a novel system of macrosteatotic hepatocytes. Methods: Hepatocytes isolated from lean Zucker rats were cultured in a collagen sandwich, incubated for 6 days in fatty acid-supplemented medium to induce steatosis, and then switched for 2 days to medium supplemented with lipid metabolism promoting agents. Intracellular lipid droplet size distribution and triglyceride, viability, albumin and urea secretion, and bile canalicular function were measured. Results: Fatty acid-supplemented medium induced microsteatosis in 3 days and macrosteatosis in 6 days, the latter evidenced by large lipid droplets dislocating the nucleus to the cell periphery. Macrosteatosis significantly impaired all functions tested. Macrosteatosis decreased upon returning hepatocytes to standard medium, and the rate of decrease was 4-fold faster with supplemented agents, yielding 80% reduction in 2 days. Viability of macrosteatosis reduced hepatocytes was similar to control lean cells. Accelerated macrosteatotic reduction led to faster recovery of urea secretion and bile canalicular function, but not of albumin secretion. Conclusions: Macrosteatosis reversibly decreases hepatocyte function and supplementary agents accelerate macrosteatosis reduction and some functional restoration with no effect on viability. This in vitro model may be useful to screen agents for macrosteatotic reduction in livers before transplantation. (C) 2013 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

【 授权许可】

Free