Alginate Microencapsulated Hepatocytes Optimised for Transplantation in Acute Liver Failure

Jitraruch, Suttiruk; Dhawan, Anil; Hughes, Robin D.; Filippi, Céline; Soong, Daniel; Philippeos, Christina; Lehec, Sharon C.; Heaton, Nigel; Longhi, Maria Serena; Mitry, Ragai R.

doi:10.1371/journal.pone.0113609

Cited by 109 publications

(103 citation statements)

References 36 publications

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“…Primary human hepatocytes cultured on alginate microbeads in vitro for 3 days showed albumin and urea production. In addition, the intraperitoneal transplantation of hepatocyte microbeads improved liver function up to 7 days in an animal model of acute liver failure 20. Despite these encouraging results, it is necessary to achieve more scientific insights and technical validation of both long‐term in vitro culture and in vivo implantation before this technology can be proposed for clinical applications.…”

Section: Implantable Technologies For Liver Therapiesmentioning

confidence: 99%

Liver tissue engineering: From implantable tissue to whole organ engineering

Mazza

Al‐Akkad

Rombouts

et al. 2017

Hepatology Communications

107

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The term “liver tissue engineering” summarizes one of the ultimate goals of modern biotechnology: the possibility of reproducing in total or in part the functions of the liver in order to treat acute or chronic liver disorders and, ultimately, create a fully functional organ to be transplanted or used as an extracorporeal device. All the technical approaches in the area of liver tissue engineering are based on allocating adult hepatocytes or stem cell‐derived hepatocyte‐like cells within a three‐dimensional structure able to ensure their survival and to maintain their functional phenotype. The hosting structure can be a construct in which hepatocytes are embedded in alginate and/or gelatin or are seeded in a pre‐arranged scaffold made with different types of biomaterials. According to a more advanced methodology termed three‐dimensional bioprinting, hepatocytes are mixed with a bio‐ink and the mixture is printed in different forms, such as tissue‐like layers or spheroids. In the last decade, efforts to engineer a cell microenvironment recapitulating the dynamic native extracellular matrix have become increasingly successful, leading to the hope of satisfying the clinical demand for tissue (or organ) repair and replacement within a reasonable timeframe. Indeed, the preclinical work performed in recent years has shown promising results, and the advancement in the biotechnology of bioreactors, ex vivo perfusion machines, and cell expansion systems associated with a better understanding of liver development and the extracellular matrix microenvironment will facilitate and expedite the translation to technical applications. (Hepatology Communications 2018;2:131–141)

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Section: Implantable Technologies For Liver Therapiesmentioning

confidence: 99%

Liver tissue engineering: From implantable tissue to whole organ engineering

Mazza

Al‐Akkad

Rombouts

et al. 2017

Hepatology Communications

107

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“…Human and rat hepatocytes were encapsulated using the optimized technique described by Jitraruch et al, 4 where the microbeads were produced at a cell density of 3.5 Â 10 6 cell/ mL alginate and polymerized for 15 min. Briefly, hepatocyte microbeads were produced using the IE-50R encapsulator (Inotech Encapsulation AG, Dottikon, Switzerland) and sterile clinical grade reagents.…”

Section: Encapsulation Of Hepatocytesmentioning

confidence: 99%

“…Viability and function of microbeads (HMBs and RMBs) were assessed according to Jitraruch et al 4 immediately postthaw (day 0), day 1, day 3, and day 7. Briefly, viability of hepatocyte microbeads was evaluated by a cell membrane integrity assay using 10-mg fluorescein diacetate (FDA; Sigma-Aldrich, Gilingham, UK) and 20-mg propidium iodide (PI; Sigma-Aldrich) incubated in the dark for 90 s then visualized under a fluorescent microscope (Leica Microsystems Ltd, Milton Keynes, UK).…”

Section: Hepatocyte Microbead Viability and Functionmentioning

confidence: 99%

“…[1][2][3] Transplantation of alginate microencapsulated hepatocytes (microbeads) has been studied in animal models of ALF with promising outcomes. [4][5][6][7] Hepatocyte microbeads can replace the missing detoxification and synthetic functions of damaged hepatocytes either for a short period, bridging the patient to liver transplantation, or allowing time for the liver to regenerate and recover, avoiding liver transplantation altogether.…”

Section: Introductionmentioning

confidence: 99%

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Cryopreservation of Hepatocyte Microbeads for Clinical Transplantation

et al. 2017

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Intraperitoneal transplantation of hepatocyte microbeads is an attractive option for the management of acute liver failure. Encapsulation of hepatocytes in alginate microbeads supports their function and prevents immune attack of the cells. Establishment of banked cryopreserved hepatocyte microbeads is important for emergency use. The aim of this study was to develop an optimized protocol for cryopreservation of hepatocyte microbeads for clinical transplantation using modified freezing solutions. Four freezing solutions with potential for clinical application were investigated. Human and rat hepatocytes cryopreserved with University of Wisconsin (UW)/10% dimethyl sulfoxide (DMSO)/5% (300 mM) glucose and CryoStor CS10 showed better postthawing cell viability, attachment, and hepatocyte functions than with histidine-tryptophan-ketoglutarate/10% DMSO/5% glucose and Bambanker. The 2 freezing solutions that gave better results were studied with human and rat hepatocytes microbeads. Similar effects on cryopreserved microbead morphology (external and ultrastructural), viability, and hepatocyte-functions post thawing were observed over 7 d in culture. UW/DMSO/glucose, as a basal freezing medium, was used to investigate the additional effects of cytoprotectants: a pan-caspase inhibitor (benzyloxycarbonyl-Val-Ala-DL-Asp-fluoromethylketone [ZVAD]), an antioxidant (desferoxamine [DFO]), and a buffering and mechanical protectant (human serum albumin [HSA]) on RMBs. ZVAD (60 mM) had a beneficial effect on cell viability that was greater than with DFO (1 mM), HSA (2%), and basal freezing medium alone. Improvements in the ultrastructure of encapsulated hepatocytes and a lower degree of cell apoptosis were observed with all 3 cytoprotectants, with ZVAD tending to provide the greatest effect. Cytochrome P450 activity was significantly higher in the 3 cytoprotectant groups than with fresh microbeads. In conclusion, developing an optimized cryopreservation protocol by adding cytoprotectants such as ZVAD could improve the outcome of cryopreserved hepatocyte microbeads for future clinical use.

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“…Regarding human hepatocytes, based on the promising results in preclinical models, a team from King's College in LondoneUK has for the first time transplanted microencapsulated human hepatocytes in a child with acute liver failure [20], with success. Recently, the group reported optimized protocols to produce GMP-grade alginate-encapsulated human hepatocytes that are suitable for clinical transplantation [65].…”

Section: Provisional Clinical Trials With a Porcine Hepatocyte Productmentioning

confidence: 99%

Current status of hepatocyte xenotransplantation

Meier

Navarro-Alvarez

Morel

et al. 2015

International Journal of Surgery

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The treatment of acute liver failure, a condition with high mortality, comprises optimal clinical care, and in severe cases liver transplantation. However, there are limitations in availability of organ donors. Hepatocyte transplantation is a promising alternative that could fill the medical need, in particular as the bridge to liver transplantation. Encapsulated porcine hepatocytes represent an unlimited source that could function as a bioreactor requiring minimal immunosuppression. Besides patients with acute liver failure, patients with alcoholic hepatitis who are unresponsive to a short course of corticosteroids are a target for hepatocyte transplantation. In this review we present an overview of the innate immune barriers in hepatocyte xenotransplantation, including the role of complement and natural antibodies; the role of phagocytic cells and ligands like CD47 in the regulation of phagocytic cells; and the role of Natural Killer cells. We present also some illustrations of physiological species incompatibilities in hepatocyte xenotransplantation, such as incompatibilities in the coagulation system. An overview of the methodology for cell microencapsulation is presented, followed by proof-of-concept studies in rodent and nonhuman primate models of fulminant liver failure: these studies document the efficacy of microencapsulated porcine hepatocytes which warrants progress towards clinical application. Lastly, we present an outline of a provisional clinical trial, that upon completion of preclinical work could start within the upcoming 2-3 years

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Alginate Microencapsulated Hepatocytes Optimised for Transplantation in Acute Liver Failure

Cited by 109 publications

References 36 publications

Liver tissue engineering: From implantable tissue to whole organ engineering

Liver tissue engineering: From implantable tissue to whole organ engineering

Cryopreservation of Hepatocyte Microbeads for Clinical Transplantation

Current status of hepatocyte xenotransplantation

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