Preparation of a three-dimensional extracellular matrix by decellularization of rabbit livers

Nari, Gustavo; Cid, Mariana Paula; Comín, Romina; Reyna, Laura; Juri, Gustavo; Taborda, Ricardo; Salvatierra, Nancy Alicia

doi:10.4321/s1130-01082013000300004

Cited by 29 publications

(27 citation statements)

References 22 publications

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“…In this case, investigators used an antegrade perfusion through the portal vein at a constant flow rate and were able to obtain a translucent acellular tissue within several days. Subsequently, several protocols have been developed to obtain nonhuman liver scaffolds 47, 48, 49, 50. The resulting 3D ECM scaffolds have been shown to provide an excellent environment for the in vitro growth of multiple liver cell types retaining excellent functionality 51, 52.…”

Section: Decellularized 3d Ecm Scaffolds For Tissue/whole Organ Enginmentioning

confidence: 99%

Liver tissue engineering: From implantable tissue to whole organ engineering

Mazza

Al‐Akkad

Rombouts

et al. 2017

Hepatology Communications

113

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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: Decellularized 3d Ecm Scaffolds For Tissue/whole Organ Enginmentioning

confidence: 99%

Liver tissue engineering: From implantable tissue to whole organ engineering

Mazza

Al‐Akkad

Rombouts

et al. 2017

Hepatology Communications

113

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“…В литературе описаны успешные эксперимен-ты по децеллюляризации целой печени животных: крыс [4], кроликов [5], свиньи [6]. Также описан эксперимент по децеллюляризации левой латераль-ной доли, взятой у пациента с гемангиомой пече-ни [7].…”

Section: Abstract: Liver Decellularization Extracellular Matrix Tunclassified

Technology features of decellularization of human liver fragments as tissue-specific fine-grained matrix for cell-engineering liver construction

Немец

Кирсанова

Басок

et al. 2018

RJTAO

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1 ФГБУ «Национальный медицинский исследовательский центр трансплантологии и искусственных органов имени академика В.И. Шумакова» Минздрава России, Москва, Российская Федерация 2 ФГАОУ ВО «Первый Московский государственный медицинский университет имени И.М. Сеченова» Минздрава России (Сеченовский университет), Москва, Российская Федерация К одной из актуальных задач при создании биоинженерной печени как альтернативы трансплантации донорской печени при лечении терминальных стадий печеночной недостаточности относится поиск матрикса, способного временно выполнять функции внеклеточного матрикса (ВКМ) печени и обеспечи-вать необходимые условия для поддержания жизнеспособности и функциональной эффективности кле-ток. Основным недостатком матриксов из резорбируемых биополимерных материалов является отсутс-твие тканеспецифичности и невозможность воспроизведения уникальной структуры ВКМ печени. Цель настоящей работы состояла в разработке технологии получения децеллюляризированных фрагментов печени с сохранением структурных свойств ее ВКМ. Материалы и методы. Децеллюляризацию меха-нически измельченных фрагментов печени человека проводили в трех сменах буферного раствора (рН = 7,4), содержащего 0,1% додецилсульфата натрия и повышающуюся концентрацию Тритона Х100 (1, 2 и 3% соответственно). Исследовали влияние длительности, условий отмывки (статика, динамика, рота-ционная система, магнитная мешалка) и способов измельчения печени на сохранность структуры ВКМ печени и полноту удаления клеточных элементов и детрита. Срезы децеллюляризованных образцов фрагментов печени окрашивали гематоксилином и эозином, а также методом Массона для выявления соединительно-тканных элементов. Результаты и обсуждение. Методами гистологического анализа по-казано, что оптимальным является режим отмывки фрагментов печени в течение 3 суток при комнатной температуре в статике, сопровождающийся перемешиванием на магнитной мешалке 2-3 раза в сутки в течение 1 ч. Более длительное время или большая кратность режима перемешивания сопровождается увеличением риска нарушения структуры печеночной ткани. Предложен алгоритм предварительного исследования донорской печени человека с целью оптимизации процесса получения децеллюляризи-рованных фрагментов печени. Выводы. Предложен алгоритм оценки пригодности донорской печени человека для децеллюляризации и найдены оптимальные условия получения децеллюляризированных фрагментов печени с сохранением структуры ВКМ печени и полным удалением клеточных элементов и детрита.

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“…However, the exclusive application of this detergent resulted in incomplete decellularization with nuclear material remaining in the scaffold. In most previous studies, a combination of these two detergents was applied [6,8,10,13,14,16,22,24,26,29,35,36]. As a result, no remaining cellular and nuclear material was detected, and the structure of the extracellular components was maintained (fig.…”

Section: The Decellularization Processmentioning

confidence: 99%

“…In fundamental studies, rodents such as mice, rats, ferrets and rabbits are used [5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24]. In translational studies, large animals such as pigs and sheep are preferred [25,26,27,28,29,30,31,32].…”

Section: Source Of Organs For Liver Engineeringmentioning

confidence: 99%

See 1 more Smart Citation

Bioengineered Livers: A New Tool for Drug Testing and a Promising Solution to Meet the Growing Demand for Donor Organs

et al. 2016

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Background: Organ engineering is a new innovative strategy to cope with two problems: the need for physiological models for pharmacological research and donor organs for transplantation. A functional scaffold is generated from explanted organs by removing all cells (decellularization) by perfusing the organ with ionic or nonionic detergents via the vascular system. Subsequently the acellular scaffold is reseeded with organ-specific cells (repopulation) to generate a functional organ. Summary: This review gives an overview of the state of the art describing the decellularization process, the subsequent quality assessment, the repopulation techniques and the functional assessment. It emphasizes the use of scaffolds as matrix for culturing human liver cells for drug testing. Further, it highlights the techniques for transplanting these engineered scaffolds in allogeneic or xenogeneic animals in order to test their biocompatibility and use as organ grafts. Key Messages: The first issue is the so-called decellularization, which is best explored and resulted in a multitude of different protocols. The most promising approach seems to be the combination of pulsatile perfusion of the liver with Triton X-100 and SDS via hepatic artery and portal vein. Widely accepted parameters of quality control include the quantitative assessment of the DNA content and the visualization of eventually remaining nuclei confirmed by HE staining. Investigations regarding the composition of the extracellular matrix focused on histological determination of laminin, collagen, fibronectin and elastin and remained qualitatively. Repopulation is the second issue which is addressed. Selection of the most suitable cell type is a highly controversial topic. Currently, the highest potential is seen for progenitor and stem cells. Cells are infused into the scaffold and cultured under static conditions or in a bioreactor allowing dynamic perfusion of the scaffold. The quality of repopulation is mainly assessed by routine histology and basic functional assays. These promising results prompted to consider the use of a liver scaffold repopulated with human cells for pharmacological research. Transplantation of the (repopulated) scaffold is the third topic which is not yet widely addressed. Few studies report the heterotopic transplantation of repopulated liver tissue without vascular anastomosis. Even fewer studies deal with the heterotopic transplantation of a scaffold or a repopulated liver lobe. However, observation time was still limited to hours, and long-term graft survival has not been reported yet. These exciting results emphasize the potential of this new and promising strategy to create physiological models for pharmacological research and to generate liver grafts for the transplant community to treat organ failure. However, the scientific need for further development in the field of liver engineering is still tremendous.

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Preparation of a three-dimensional extracellular matrix by decellularization of rabbit livers

Abstract: the proposed decellularization protocol was correct, and was verified by an absence of cells. The hepatic matrix had preserved vascular and bile ducts with a suitable three-dimensional architecture permitting further cell seeding.

Cited by 29 publications

References 22 publications

Liver tissue engineering: From implantable tissue to whole organ engineering

Liver tissue engineering: From implantable tissue to whole organ engineering

Technology features of decellularization of human liver fragments as tissue-specific fine-grained matrix for cell-engineering liver construction

Bioengineered Livers: A New Tool for Drug Testing and a Promising Solution to Meet the Growing Demand for Donor Organs

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