Acellular liver matrix improves the survival and functions of isolated rat hepatocytes cultured in vitro.

Burra, P.; Tomat, S.; Conconi, Maria Teresa; Macchi, C; Russo, Francesco Paolo; Parnigotto, Pier Paolo; Naccarato, R.; Nussdorfer, Gastone G.

doi:10.3892/ijmm.14.4.511

Cited by 17 publications

(16 citation statements)

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“…An in vitro study using DLM for hepatocyte cultures has been reported 21. It was shown that human hepatocytes cultured between 2 layers of porcine liver decellularized matrix in vitro for 10 days exhibited liver‐specific functions similar to those of cells grown in a Matrigel sandwich,21 and rat hepatocytes seeded between sheets of DLM showed good viability and function in vitro 22, 23. Some of these previous studies employed DLM pieces, and the decellularized matrix tissue was lyophilized into a powder form and was rehydrated to generate a gel‐like carrier.…”

Section: Discussionmentioning

confidence: 99%

Decellularized liver matrix as a carrier for the transplantation of human fetal and primary hepatocytes in mice

et al. 2011

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The transplantation of primary hepatocytes has been shown to augment the function of damaged livers and to bridge patients to liver transplantation. However, primary hepatocytes often have low levels of engraftment and survive for only a short time after transplantation. To explore the potential benefits of using decellularized liver matrix (DLM) as a carrier for hepatocyte transplantation, DLM from whole mouse livers was generated. Human fetal hepatocytes immortalized by telomerase reconstitution (FH-hTERTs) or primary human hepatocytes were infused into the DLM, which was then implanted into the omenta of immunodeficient nonobese diabetic/severe combined immunodeficient/interleukin-2 receptor c-deficient mice or nonobese diabetic/severe combined immunodeficient/mucopolysaccharidosis type VII mice. The removal of endogenous cellular components and the preservation of the extracellular matrix proteins and vasculature were demonstrated in the resulting DLM. Bioluminescent imaging revealed that FH-hTERTs transduced with a lentiviral vector expressing firefly luciferase survived in the DLM for 8 weeks after peritoneal implantation, whereas the luciferase signal from FH-hTERTs rapidly declined in control mice 3 to 4 weeks after transplantation via splenic injection or omental implantation after Matrigel encapsulation. Furthermore, primary human hepatocytes that were reconstituted in the DLM not only survived 6 weeks after transplantation but also maintained their function, as demonstrated by messenger RNA levels of albumin and cytochrome P450 (CYP) subtypes (CYP3A4, CYP2C9, and CYP1A1) similar to the levels in freshly isolated human primary hepatocytes (hPHs). In contrast, when hPHs were transplanted into mice via splenic injection, they failed to express CYP3A4, although they expressed albumin. In conclusion, DLM provides an excellent environment for long-term survival and maintenance of the hepatocyte phenotype after transplantation.

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Section: Discussionmentioning

confidence: 99%

Decellularized liver matrix as a carrier for the transplantation of human fetal and primary hepatocytes in mice

et al. 2011

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“…Moreover, studies on ß-cells cultured on various crude matrices or on their purified extracts highlighted the (17,(21)(22)(23)(24)(25)(26). Tissue engineering studies have also shown that scaffolds derived from decellularized matrices, obtained after the removal of the cellular component, support in vitro adhesion, growth, and function of several cell types (27)(28)(29) and act as a template for growth and tissue remodelling (30)(31)(32)(33)(34)(35). Based on these considerations, in this study we ascertained whether pancreas and/or liver acellular matrices support the pancreatic islet secretory function.…”

Section: Introductionmentioning

confidence: 99%

Pancreatic acellular matrix supports islet survival and function in a synthetic tubular device: In vitro and in vivo studies

Carlo

Baiguera²,

Conconi³

et al. 2009

Int J Mol Med

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Abstract. Increasing pancreatic islet survival and function is a starting point for obtaining a valuable bioartificial pancreas for the treatment of type 1 diabetes. In this context, decellularized matrices, obtained after the removal of tissue cellular part, are known to support in vitro adhesion, growth, and function of several cell types. We demonstrate that a homologous acellular pancreatic matrix is a suitable scaffold for rat islet cultures maintaining their long-term viability and function. Islets adhered to the pancreatic matrix showed a constant glucose-induced insulin release during long-term in vitro incubation, while islets cultured without a matrix or on the liver matrix showed a progressive reduction. In order to obtain implantable devices, acellular matrix/islet cultures were entrapped into poly(vinyl alcohol) (PVA)/ poly(ethylene glycol) (PEG) tubes obtained by the freezing/thawing procedure. Under this condition, an in vitro constant insulin release was detected. The devices were then implanted into diabetic rats where reduced insulin requirement was noted suggesting insulin secretory activity of islets contained in the device. Indeed, immunofluorescence confirmed the presence of insulin-and glucagon-producing cells into the explanted devices. These data show that PVA/PEG semi-permeable membrane can obtain devices that restore, at least in part, insulin secretion.

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“…However, the current study has several limitations and some points to be investigated. In this study, the presence of growth factors in ECM of the scaffolds were not verified which are necessary to determine the adhesion, migration, differentiation, proliferation, and survival of seeded cells . Furthermore, no access to blood vessels was made during the surgery, which may affect the in vivo recellularization process.…”

Section: Discussionmentioning

confidence: 97%

Whole-organ tissue engineering: Decellularization and recellularization of three-dimensional matrix liver scaffolds

Sabetkish

Kajbafzadeh

Sabetkish

et al. 2014

J. Biomed. Mater. Res.

133

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To report the results of whole liver decellularization by two different methods. To present the results of grafting rat and sheep decellularized liver matrix (DLM) into the normal rat liver and compare natural cell seeding process in homo/xenograft of DLM. To compare the results of in vitro whole liver recellularization with rats' neonatal green fluorescent protein (GFP)-positive hepatic cells with outcomes of in vivo recellularization process. Whole liver of 8 rats and 4 sheep were resected and cannulated via the hepatic vein and perfused with sodium dodecyl sulfate (SDS) or Triton + SDS. Several examinations were performed to compare the efficacy of these two decellularization procedures. In vivo recellularization of sheep and rat DLMs was performed following transplantation of multiple pieces of both scaffolds in the subhepatic area of four rats. To compare the efficacy of different scaffolds in autologous cell seeding, biopsies of homograft and xenograft were assessed 8 weeks postoperatively. Whole DLMs of 4 rats were also recellularized in vitro by perfusion of rat's fetal GFP-positive hepatic cells with pulsatile bioreactor. Histological evaluation and enzymatic assay were performed for both in vivo and in vitro recellularized samples. The results of this study demonstrated that the triton method was a promising decellularization approach for preserving the three-dimensional structure of liver. In vitro recellularized DLMs were more similar to natural ones compared with in vivo recellularized livers. However, homografts showed better characteristics with more organized structure compared with xenografts. In vitro recellularization of liver scaffolds with autologous cells represents an attractive prospective for regeneration of liver as one of the most compound organs. In vivo cell seeding on the scaffold of the same species may have more satisfactory outcomes when compared with the results of xenotransplantation. This study theoretically may pave the road for in situ liver regeneration probably by implantation of homologous DLM or in vitro recellularized scaffolds into the diseased host liver.

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Acellular liver matrix improves the survival and functions of isolated rat hepatocytes cultured in vitro.

Cited by 17 publications

References 0 publications

Decellularized liver matrix as a carrier for the transplantation of human fetal and primary hepatocytes in mice

Decellularized liver matrix as a carrier for the transplantation of human fetal and primary hepatocytes in mice

Pancreatic acellular matrix supports islet survival and function in a synthetic tubular device: In vitro and in vivo studies

Whole-organ tissue engineering: Decellularization and recellularization of three-dimensional matrix liver scaffolds

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