Method for the decellularization of intact rat liver

Shupe, Thomas; Williams, Matthew J.; Brown, Alicia; Willenberg, Bradley J.; Petersen, Bryon E.

doi:10.4161/org.6.2.11546

Cited by 155 publications

(126 citation statements)

References 9 publications

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“…These have included injecting cells directly into the parenchymal space, perfusion through the portal vein and perfusion through the hepatic vein. 13,19,24,25,27 One method which has not yet been documented is perfusion through the bile duct. The unique architecture of the portal triad suggests that the biliary infusion will theoretically be able to deliver cells throughout the liver ECM.…”

Section: Discussionmentioning

confidence: 99%

“…A number of methods have been proposed using different combinations and varying concentrations of SDS and Triton X-100 on different animal models including murine, rabbit, porcine, ferret and rat. 8,10,13,[23][24][25][26][27][28][29] The goal of this process is to remove all cellular material while retaining the native ECM structure, and most established protocols have successfully demonstrated this to be quite possible. It remains to be seen whether the preservation of growth factors and essential proteins found on the ECM will play an important role in promoting the proliferation and function of new parenchymal and nonparenchymal cells.…”

Section: Discussionmentioning

confidence: 99%

“…20 Several studies have demonstrated success with decellularization by using a lower concentration of SDS and several concentrations of Triton X-100. 25,26 We simplified the decellularization process by only using 0.1% SDS. The effectiveness of this method in removing all cellular material from the ECM was verified by H&E and DNA extraction.…”

Section: Discussionmentioning

confidence: 99%

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Recellularization via the bile duct supports functional allogenic and xenogenic cell growth on a decellularized rat liver scaffold

et al. 2016

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ABSTRACT. Recent years have seen a proliferation of methods leading to successful organ decellularization. In this experiment we examine the feasibility of a decellularized liver construct to support growth of functional multilineage cells. Bio-chamber systems were used to perfuse adult rat livers with 0.1% SDS for 24 hours yielding decellularized liver scaffolds. Initially, we recellularized liver scaffolds using a human tumor cell line (HepG2, introduced via the bile duct). Subsequent studies were performed using either human tumor cells co-cultured with human umbilical vein endothelial cells (HUVECs, introduced via the portal vein) or rat neonatal cell slurry (introduced via the bile duct). Bio-chambers were used to circulate oxygenated growth medium via the portal vein at cholangiocytes was verified by CK-7 positivity. Quantitative albumin measurement from the grafts showed increasing albumin levels after seven days of perfusion. Graft albumin production was higher than that observed in traditional cell culture. This data shows that rat liver scaffolds support human cell ingrowth. The scaffold likewise supported the engraftment and survival of neonatal rat liver cell slurry. Recellularization of liver scaffolds thus presents a promising model for functional liver engineering.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Recellularization via the bile duct supports functional allogenic and xenogenic cell growth on a decellularized rat liver scaffold

et al. 2016

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“…The use of ionic and non-ionic detergents may in turn cause damage to DNA -proteins interactions, lipids, lipoproteins, protein denaturation. It can also change the ultrastructure of the ECM [15]. Furthermore, acellularization factors may exhibit cytotoxic activity.…”

Section: Discussionmentioning

confidence: 99%

Biocompatibility Issue Of Tissue Engineered Heart Valves

Wilczek¹

2015

Archives of Metallurgy and Materials

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Tissue engineering is a new field of knowledge which creates the possibilities for producing bioactive cardiac prostheses that will characterize by biomechanical and morphological properties similar to native tissue. It is expected that it will be characterized by high durability, which is very important from the social and clinical point of view. The aim of the study was to compare the cytotoxic effect of enzymatic and detergent acellularization methods commonly used for the biological scaffold preparation. It seems that the use of enzymatic methods, allows efficient donor cells removal while maintaining the ability to autologous cell seeding. Heart valves bioprosthesis created using these techniques, may be a good alternative to the currently used prostheses.Keywords: tissue engineering, heart valve, cytotoxicity, scaffold.Inżynieria tkankowa jest nową dziedziną wiedzy, która stwarza możliwości wytwarzania protez serca bioaktywnych charakteryzujących się właściwościami biomechanicznymi i morfologicznymi zbliżonymi do tkanki rodzimej. Oczekuje się, że proteza serca będzie charakteryzować się wysoką trwałością, co jest bardzo ważne z punktu widzenia społecznego i klinicznego. Celem badania było porównanie efektu cytotoksycznego enzymatycznych i detergentowych metod usuwania komórek, powszechnie stosowanych do wytwarzania biologicznych rusztowań. Wydaje się, że stosowanie metod enzymatycznych, umożliwia wydajne usunięcie komórek dawcy przy zachowaniu zdolności do autologicznego posiewu komórek. Bioprotezy zastawek serca tworzone za pomocą tej techniki mogą być dobrą alternatywą dla obecnie stosowanych mechanicznych i biologicznych protez zastawek serca.

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“…Since we are close to reaching that goal, the article on methodology by de Kock et al 2011 cannot be valued high enough in embarking on a novel approach to hepatic function analysis. The method of decellularization of intact rat liver (Shupe et al 2010;Bao et al 2010) is providing an intact decellularized organ with an intact liver capsule and all applications looked for; furthermore, re-cellularization of the tissue engineered liver graft enables liver-speciWc functions such as albumin secretion, urea synthesis, and cytochrome P 450 expression at comparable levels of "in vivo liver" (Uygun et al 2010). Besides of the obvious potential for the generation of a transplantable liver graft as therapeutic principle for liver disease, this technical approach holds potential for in vitro applications in toxicology, pharmacology and alternative methods versus in vivo testing in laboratory animals.…”

mentioning

confidence: 99%

Good news for sound science-based alternatives in liver

Garthoff¹

2011

Arch Toxicol

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Method for the decellularization of intact rat liver

Cited by 155 publications

References 9 publications

Recellularization via the bile duct supports functional allogenic and xenogenic cell growth on a decellularized rat liver scaffold

Recellularization via the bile duct supports functional allogenic and xenogenic cell growth on a decellularized rat liver scaffold

Biocompatibility Issue Of Tissue Engineered Heart Valves

Good news for sound science-based alternatives in liver

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