Clinical translation of bioartificial liver support systems with human pluripotent stem cell-derived hepatic cells

Sakiyama, Ryoichi; Blau, Brandon; Miki, Toshio

doi:10.3748/wjg.v23.i11.1974

Cited by 37 publications

(22 citation statements)

References 38 publications

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“…At present no BAL is in use to clinically treat liver failure. Some clinical trials have been conducted (for an updated review consult the work of Sakiyama et al[ 30 ] and references therein) with some success. All of these devices use isolated liver-derived cells.…”

Section: Discussionmentioning

confidence: 99%

Experimental bio-artificial liver: Importance of the architectural design on ammonia detoxification performance

Pizarro¹,

Mamprin²,

Daurelio³

et al. 2018

WJH

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AIMTo determine the influence of the construction design over the biological component’s performance in an experimental bio-artificial liver (BAL) device.METHODSTwo BAL models for liver microorgans (LMOs) were constructed. First, we constructed a cylindrical BAL and tested it without the biological component to establish its correct functioning. Samples of blood and biological compartment (BC) fluid were taken after 0, 60, and 120 min of perfusion. Osmolality, hematocrit, ammonia and glucose concentrations, lactate dehydrogenase (LDH) release (as a LMO viability parameter), and oxygen consumption and ammonia metabolizing capacity (as LMO functionality parameters) were determined. CPSI and OTC gene expression and function were measured. The second BAL, a “flat bottom” model, was constructed using a 25 cm2 culture flask while maintaining all other components between the models. The BC of both BALs had the same capacity (approximately 50 cm3) and both were manipulated with the same perfusion system. The performances of the two BALs were compared to show the influence of architecture.RESULTSThe cylindrical BAL showed a good exchange of fluids and metabolites between blood and the BC, reflected by the matching of osmolalities, and glucose and ammonia concentration ratios after 120 min of perfusion. No hemoconcentration was detected, the hematocrit levels remained stable during the whole study, and the minimal percentage of hemolysis (0.65% ± 0.10%) observed was due to the action of the peristaltic pump. When LMOs were used as biological component of this BAL they showed similar values to the ones obtained in a Normothermic Reoxygenation System (NRS) for almost all the parameters assayed. After 120 min, the results obtained were: LDH release (%): 14.7 ± 3.1 in the BAL and 15.5 ± 3.2 in the NRS (n = 6); oxygen consumption (μmol/min·g wet tissue): 1.16 ± 0.21 in the BAL and 0.84 ± 0.15 in the NRS (n = 6); relative expression of Cps1 and Otc: 0.63 ± 0.12 and 0.67 ± 0.20, respectively, in the BAL, and 0.86 ± 0.10 and 0.82 ± 0.07, respectively, in the NRS (n = 3); enzymatic activity of CPSI and OTC (U/g wet tissue): 3.03 ± 0.86 and 222.0 ± 23.5, respectively, in the BAL, and 3.12 ± 0.73 and 228.8 ± 32.8, respectively, in the NRS (n = 3). In spite of these similarities, LMOs as a biological component of the cylindrical BAL were not able to detoxify ammonia at a significant level (not detected vs 35.1% ± 7.0% of the initial 1 mM NH4+ dose in NRS, n = 6). Therefore, we built a second BAL with an entirely different design that offers a flat base BC. When LMOs were placed in this “flat bottom” device they were able to detoxify 49.3% ± 8.8% of the initial ammonia overload after 120 min of perfusion (n = 6), with a detoxification capacity of 13.2 ± 2.2 μmol/g wet tissue.CONCLUSIONIn this work, we demonstrate the importance of adapting the BAL architecture to the biological component characteristics to obtain an adequate BAL performance.

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Section: Discussionmentioning

confidence: 99%

Experimental bio-artificial liver: Importance of the architectural design on ammonia detoxification performance

Pizarro¹,

Mamprin²,

Daurelio³

et al. 2018

WJH

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“…Bioartificial liver (BAL) is an extracorporeal bioarti-ficial organ device based on functional liver cell culture, which is able to temporarily replace liver detoxification, synthesis, secretion and conversion among other functions. Patients who receive BAL treatment may have an increased survival rate while waiting for liver transplantation and may also have improved liver regeneration ( 4 – 6 ).…”

Section: Introductionmentioning

confidence: 99%

Reversibly immortalized hepatic progenitor cell line containing double suicide genes

Fang

Liu

et al. 2018

Int J Mol Med

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A large number of functional hepatocytes is required for bioartificial liver therapy. Simian virus 40 T-antigen (SV40T) has been previously reported to improve the immortalized proliferation of primary hepatocytes to generate a sufficient number of cells; however, these long-term immortalized hepatocytes may induce further malignant transformation in vivo. In the present study, the SV40T immortalization gene and two suicide genes, herpes simplex virus thymidine kinase (HSV-tk) and cytosine deaminase (CD), were transducted into primary hepatocytes to construct a novel type of Cre/LoxP-mediated reversible immortalized hepatocyte line. Polymerase chain reaction analysis and western blotting confirmed that the SV40T, HSV-tk and CD genes were successfully inserted into hepatic progenitor cells and their expression was controlled by Cre/LoxP recombination. Total removal of SV40T could be achieved via the ganciclovir (GCV)/HSV-tk suicide system. Cells maintained their biosafety in vivo with CD gene expression and 5-fluoro-cytosine (5-FC) induced cell death. Following transplantation into the carbon tetrachloride (CCl4) model group, the majority of cells had survived after 14 days post-implantation and a number of the cells had transported into the liver parenchyma. When compared with the CCl4 model group, the transplanted cells repaired the liver biochemical index and pathological structure markedly. Thus, the present study reports a novel reversible immortalized hepatocyte with double suicide genes, which exhibited the cellular phenotype and recovery function of normal liver cells. This method maximally guaranteed the biological safety of immortalized hepatocytes for in vivo application, providing a reliable, safe and ideal cell material for the artificial liver technique.

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“…Although the use of iPSCs was successful in the reproduction of the phenotypes of those diseases, its application for pancreatic diseases, such as pancreatitis and diabetes (Borowiak and Melton, 2009), and some other diseases of the kidney (Mae and Osafune, 2015) and liver (Sakiyama et al, 2017) remains to be successfully performed, presumably due to the difficulty of cell differentiation in vivo and the complexity of the organ structures. In order to reproduce the diseases that are derived from tissues, in which induction of differentiation is difficult, it is undoubtedly necessary to improve the efficiency of differentiation induction and to reconstruct the complexity of tissues in a model.…”

Section: Introductionmentioning

confidence: 99%

Hereditary Pancreatitis Model by Blastocyst Complementation in Mouse

Asai

Konno²,

Koseki

et al. 2018

Preprint

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StatementThe present study is the first report for reproducing human disease by utilizing blastocyst complementation method and would lead to the development of novel therapy for human disease. AbstractThe application of pluripotent stem cell is expected to contribute to the elucidation of the unknown mechanism of human diseases. However, in vitro induction of cells in several organs, such as the pancreas and liver, remains difficult; therefore, reproduction of those diseases in a model has not been feasible. To reproduce human hereditary pancreatitis (HP), which is most frequently caused by the mutations in the PRSS1 gene, we performed the blastocyst complementation (BC) method. In the BC method, mouse embryonic stem (ES) cells harboring CRISPR/CAS9-mediated mutations in the Prss1 were injected into the blastocysts of deficient Pdx1 gene mice, which is a critical transcription factor in the pancreas. The results showed that the blastocysts injected into the Prss1-mutant ES cells induced trypsin activation. This implied that the mouse phenotype mimics that of human HP and that the BC method was useful for the reproduction and study of pancreatic disorders. The present study opens the possibility of investigating uncharacterized human diseases by utilizing the BC method.

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Clinical translation of bioartificial liver support systems with human pluripotent stem cell-derived hepatic cells

Cited by 37 publications

References 38 publications

Experimental bio-artificial liver: Importance of the architectural design on ammonia detoxification performance

Experimental bio-artificial liver: Importance of the architectural design on ammonia detoxification performance

Reversibly immortalized hepatic progenitor cell line containing double suicide genes

Hereditary Pancreatitis Model by Blastocyst Complementation in Mouse

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