Date: Saturday, May 30, 2020
Session Time: 3:15pm-4:00pm
Presentation Time: 3:30pm-4:00pm
*Purpose: Acute liver failure (ALF) has a rapid and unpredictable onset with high mortality. The only current treatment is to transplant a donor organ but donor organ shortages exist. A bioartificial liver (BAL) could bridge the gap to transplant or buy time for spontaneous recovery. In order to treat ALF in a timely fashion, a hepatocyte bank system capable of supplying large numbers of hepatocytes must be established. However, hepatocytes are very susceptible to cryopreservation injury. On thawing, cryopreserved hepatocytes often have reduced viability and metabolic function in comparison with fresh cells. We developed an optimized protocol for cryopreservation of hepatocyte spheroids using serum-free cryopreservation solution. We evaluated the efficacy of BAL system, containing cryopreserved porcine hepatocytes spheroids, in a porcine model of ALF.
*Methods: Porcine hepatocytes with a viability of 85~96% were isolated from male pigs weighing 14 to 18 kg, raised in a sterile environment. The isolate hepatocytes were cultured to form spheroids and were frozen without serum in liquid nitrogen by the use of a controlled rate Freezer. After thawing, hepatocyte spheroids were mixed with 1.5% alginate solution and placed in a high content/speed immobilization apparatus and dropped into 100mM calcium solution. The Ca-alginate-immobilized hepatocyte spheroids beads were packed within the bioreactor of the BAL system. BAL treatment was continued for up to 12 hours.
*Results: ALF pigs were divided into three groups. The control group consisted of treatment naïve pigs (n = 5), blank group consisted of pigs that were attached to the BAL system not containing hepatocytes for 12 hours (n = 5) and BAL group consisted of pigs that were attached to the BAL containing hepatocytes for 12 hours (n = 5). Increase in serum ammonia levels were significantly greater in the blank group (P < 0.05) and control group (P < 0.05), compared to the BAL group during the treatment period. Survival was significantly prolonged in the BAL group compared to the blank group (P =0.027) and control group (P =0.004).
*Conclusions: A BAL system with a bioreactor containing hepatocyte spheroids showed effective clearance of serum ammonia, preservation of renal function and prolonged survival in a porcine model of ALF.
To cite this abstract in AMA style:Lee S, Lee J, Park H, Kim Y, Noh J, Jung J, Lee S, Shin H, Yoon H, Lee D, Lee S. Improved Survival of Acute Liver Failure by “Off-the-Shelf” Bioartificial Liver Support System Using Immobilized Cryopreserved Hepatocyte Spheroids in Porcine Model [abstract]. Am J Transplant. 2020; 20 (suppl 3). https://atcmeetingabstracts.com/abstract/improved-survival-of-acute-liver-failure-by-off-the-shelf-bioartificial-liver-support-system-using-immobilized-cryopreserved-hepatocyte-spheroids-in-porcine-model/. Accessed February 28, 2021.
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