Optimizing Perfusion-Decellularization Methods of Porcine Livers for Clinical-Scale Whole-Organ Bioengineering

Wu, Qiong; Bao, Ji; Zhou, Yongjie; Wang, Yujia; Du, Zhenggui; Shi, Yujun; Li, Li; Bu, Hong

doi:10.1155/2015/785474

Cited by 46 publications

(85 citation statements)

References 34 publications

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“…Tchen et al [2007] reported that detergents like SDS/ Triton X-100 would disrupt the ECM ultrastructure in pancreatic bioscaffold with a high fat content. SDS tends to destroy the native tissue ultrastructure and can destabilize the triple-helical domain and integrity of collagen [Wu et al, 2015b]. In this study, a colorimetric assay was used to detect hydroxyproline, a major element of collagen.…”

Section: Discussionmentioning

confidence: 99%

Decellularized Pancreas Matrix Scaffolds for Tissue Engineering Using Ductal or Arterial Catheterization

Hashemi

Pasalar

Soleimani

et al. 2018

Cells Tissues Organs

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Introduction: Diabetes is known as a worldwide disease with a great burden on society. Since therapeutic options cover a limited number of target points, new therapeutic strategies in the field of regenerative medicine are considered. Bioscaffolds along with islet cells would provide bioengineered tissue as a substitute for β-cells. The perfusion-decellularization technique is considered to create such scaffolds since they mimic the compositional, architectural, and biomechanical nature of a native organ. In this study, we investigated 2 decellularization methods preserving tissue microarchitecture. Methods: Procured pancreas from Sprague-Dawley rats was exposed to different percentages of detergent for 2, 4, and 6 h after cannulation via the common bile duct or aorta. Results: High concentrations of sodium dodecyl sulfate (SDS), i.e., > 0.05%, resulted in tissue disruption or incomplete cell removal depending on the duration of exposure. In both methods, 6-h exposure to 0.05% SDS created a bioscaffold with intact extracellular matrices and proper biomechanical characteristics. Tissue-specific stainings revealed that elastic, reticular, and collagen fiber concentrations were well preserved. Quantitative findings showed that glycosaminoglycan content was slightly different, but hydroxyproline was in the range of native pancreas tissue. Dye infusion through ductal and vascular cannulation proved that the vascular network was intact, and scanning electron microscopy indicated a homogeneous porous structure. Conclusions: Using the detergent-based method, an effective and time-efficient procedure, a whole pancreas extracellular matrix bioscaffold can be developed that can be used as a 3D structure for pancreas tissue engineering-based studies and regenerative medicine applications.

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Section: Discussionmentioning

confidence: 99%

Decellularized Pancreas Matrix Scaffolds for Tissue Engineering Using Ductal or Arterial Catheterization

Hashemi

Pasalar

Soleimani

et al. 2018

Cells Tissues Organs

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“…While it has been shown that triton x‐100 is not as effective as sodium dodecyl sulfate (SDS) for DNA removal, it better preserves the matrix architecture and endogenous growth factors, which play important roles in modulating cellular functions . Studies have also demonstrated that triton‐x100 treated decellularized liver scaffolds supported superior hepatic cell functions when compared to SDS treated counterparts . Triton x‐100 has also been used for preparing liver scaffolds of a clinical relevant scale …”

Section: Discussionmentioning

confidence: 99%

Vasculature reconstruction of decellularized liver scaffolds via gelatin‐based re‐endothelialization

Meng

Almohanna

Altuhami

et al. 2018

J Biomedical Materials Res

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Decellularized liver scaffolds based liver engineering is a promising approach toward developing functional liver surrogates. However, a major obstacle to long‐term transplantation is the hemocompatibility of the bioengineered liver surrogates. One approach to improve the hemocompatibility of engineered liver surrogates is re‐endothelialization. In the current study, immortalized endothelial cells were perfused for re‐endothelialization of decellularized rat liver scaffolds. When compared to the media‐based perfusion approach, gelatin hydrogels‐based perfusion significantly increased the number of cells that were retained in the decellularized liver scaffolds and the vascular lumen coverage ratio. Endothelial cells were lining along the vasculatures of the decellularized liver scaffolds and actively proliferating. Re‐endothelialization improved the blood retention ability of the liver scaffold vasculatures. Doppler ultrasound detected active blood flows within the re‐endothelialized liver scaffold transplants 8 days post‐transplantation. Our results strengthened the feasibility of developing bioengineered liver surrogates utilizing decellularized liver scaffolds. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 392–402, 2019.

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“…Primary rat hepatocytes were isolated from male Sprague-Dawley rats (body weight, 180–220 g) by a two-step collagenase perfusion method, and their viability was confirmed to be >95% by the trypan blue exclusion method (12). Hepatocytes were cultured in serum-free medium as previously described (13).…”

Section: Methodsmentioning

confidence: 99%

Construction of a general albumin promoter reporter system for real-time monitoring of the differentiation status of functional hepatocytes from stem cells in mouse, rat and human

Tang

et al. 2017

Biomedical Reports

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Genetic constructs with promoters fused to reporter genes for simultaneous monitoring of cellular events have been the focus of attention in recent years. Adenoviral vectors, which have distinctive characteristics, have been used to monitor the differentiation of stem cells in vitro. In the present study, a modified adenoviral vector was constructed, containing a mouse, rat, and human general albumin promoter sequence fused to a ZsGreen reporter gene, and evaluated its efficiency in different cell types. Two hepatocyte cell lines (Hepa1-6 and HepG2), rat primary hepatocytes, rat bone marrow mesenchymal stem cells (BM-MSCs) and rat BM-MSCs-derived hepatocyte-like cells were transduced with this vector, and the transfection efficiency and functional capabilities of the promoter were evaluated by fluorescent microscopy. The results demonstrated efficient expression of ZsGreen in Hepa1-6 cells, HepG2 cells, rat primary hepatocytes, and rat BM-MSCs-derived hepatocyte-like cells, but not in rat BM-MSCs. In conclusion, the current study demonstrates a simple, high-efficiency, general tool for real-time monitoring of the differentiation status of hepatocytes from stem cells in mice, rats, and humans. This tool may be useful for evaluating different protocols to generate functional hepatocytes from stem cells in multiple species.

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Optimizing Perfusion-Decellularization Methods of Porcine Livers for Clinical-Scale Whole-Organ Bioengineering

Cited by 46 publications

References 34 publications

Decellularized Pancreas Matrix Scaffolds for Tissue Engineering Using Ductal or Arterial Catheterization

Decellularized Pancreas Matrix Scaffolds for Tissue Engineering Using Ductal or Arterial Catheterization

Vasculature reconstruction of decellularized liver scaffolds via gelatin‐based re‐endothelialization

Construction of a general albumin promoter reporter system for real-time monitoring of the differentiation status of functional hepatocytes from stem cells in mouse, rat and human

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