Perfusion decellularization is a novel technique to produce whole liver scaffolds that retains the organ s extracellular matrix composition and microarchitecture. Decellularization for det treatment, the pv was connected to a peristaltic pump masterflex, uk and perfused with dh 2 o 18. Enabling technology for whole organ decellularization. Akinori hirano, mio kasai, yuta akamatsu, hidetoshi oka, yuko kitagawa, hideyuki shimizu, heterotopic transplantation of a decellularized and recellularized whole porcine heart, interactive. It also proved highly beneficial in saving time as the decellularization procedure was reduced from days down to just 24 h. The optimized 3% sds decellularization method was used for these hearts at a flow rate of. Matrix decoded a pancreatic extracellular matrix with. Digital angiography videos were obtained for each injection route. Biological matrices derived by organ decellularization are of increasing significance for te strategies as they allow the recreation of the ecm microenvironment and provide important cues for cellular function. Important advances have occurred in organ bioengineering using decellularized scaffolds in small animal models. Our decellularization protocol began with the disruption of the cell membrane using freezethaw cycles. An overview of tissue and whole organ decellularization processes. An overview of tissue and whole organ decellularization. However, the development of human size whole organ decellularization to engineer liver grafts for transplantation is necessary.
Article information, pdf download for successful decellularization of. However, the function exhibited by bioengineered organs has been rudimentary. We show a method for the dissection of an intact porcine pancreas and the process of successful decellularization by perfusion of detergents triton x100, sodium deoxycholate, and deoxyribonuclease. Decellularized liver matrix at the end of the decellularization process. Perfusion decellularization is the most consistent method to obtain decellularized whole organ scaffolds to serve as a platform for organ bioengineering. Isolation and decellularization of a whole porcine. This study aimed at developing a procedure for decellularizing heart tissue harvested from rats and evaluating decellularization efficiency in terms of residual nuclear content and structural properties. Tissue engineering of the whole pancreas is a challenge because of its exocrine and endocrine functions. Decellularization methods of porcine kidneys for whole. Decellularized human liver as a natural 3dscaffold for liver bioengineering and transplantation.
Organ transplantation is now a wellestablished procedure for the treatment of endstage organ failure due to various causes, but is a victim of. An overview of tissue and whole organ decellularisation. Automated decellularization of intact, humansized lungs. Decellularization and recellularization technologies in. While the experimental studies involving the use of decellularized organ scaffolds are encouraging, the translation of whole organ. Decellularization and recellularization of threedimensional matrix scaffolds. Humanizing miniature hearts through 4flow cannulation perfusion. Decellularization and recellularization methodology for. Decellularized human liver as a natural 3dscaffold for. The decellularization process involved the use of 33cholamidopropyl dimethylammonio1propanesulfonate chaps and sodium dodecyl sulfate sds buffers for 6 h each, followed by incubation in a serum medium. By utilizing intrinsic vasculature, complex whole organ decellularization can be achieved via antegrade or retrograde perfusion 4 to remove cellular components while preserving the delicate 3d extracellular matrix and vasculature 2, 5, 6, 7. Decellularization for whole organ bioengineering iopscience.
Nonhuman primate lung decellularization and recellularization using a specialized large organ bioreactor. Whole organ decellularization a tool for bioscaffold fabrication and organ bioengineering. Decellularization is the process of removing the dna and cells from a donated organ. Repopulation of decellularized whole organ scaffold using. Optimized protocol for whole organ decellularization european. Once the scaffold is in place, the organ can be regrown with target cells. Researchers have created a beating heart in the laboratory. Decellularization for the different plant types was adapted from whole organ perfusion decellularization techniques 6. Sage video streaming video collections opens in new tab. Perfusion decellularization is the most consistent method to obtain decellularized whole organ scaffolds due to the native vascular architecture, which is naturally designed to permit the delivery.
Whole organ decellularization is a process which allows the removal of cells while leaving the chemical and structural aspects of. Comparison of methods for wholeorgan decellularization in tissue. Perfusion decellularization of whole organs request pdf. The decellularization techniques have led to the development of scaffolds for multiple organs, including the heart, liver, lung and kidney. Decellularization also spelt decellularisation in british english is the process used in biomedical engineering to isolate the extracellular matrix ecm of a tissue from its inhabiting cells, leaving an ecm scaffold of the original tissue, which can be used in artificial organ and tissue regeneration. Standardized decellularization of fetal and adult hearts permits comparative experimental studies of both tissues in a 3d context by capturing native architecture and biomechanical properties. Humanizing miniature hearts through 4flow cannulation. To summarize, this work has demonstrated that preserving the vascular accessibility with the 4flow cannulation improves whole organ perfusion reducing the amount of time needed for decellularization.
Decellularization for the different plant types was adapted from whole organ perfusion decellularization techniques. Repopulation of whole decellularized organ scaffolds using stem cells has added a new dimension for creating new bioengineered organs. Sage knowledge the ultimate social sciences library. Successful decellularization of thickwalled tissue. Scientists develop technique ensuring 100% success rate of organ transplants. A recent innovation in decellularization is the generation of wholeorgan ecm. Spinach leaves were cannulated through the petiole and parsley stems were cannulated via the basipetal end of the stem segment. We found that the combination of freezethaw cycles, 1% tx and 0. Rodriguez,1,4 anthony atala,1 and shay soker1 a major roadblock to successful organ bioengineering is the need for a functional vascular. Attaining a wellqualified whole decellularized organ applicable for an enduring and successful transplantation, decellularization protocols should be organ specific in terms of decellularizing agents and time of tissue exposure. Efficient decellularization for bovine pericardium with. Decellularization of whole urethras for tissue engineering applications.
In this study, we generated a whole organ porcine pancreasspecific ecm with conserved micro and ultrastructural features. Herein, the method of preparing whole organ scaffolds using perfusion decellularization and subsequent repopulation with hepatocytes is described. The swanson school of engineering in partial fulfillment. Inspired by previous work on whole organ decellularization 14,15,24, the main goal of this work was to establish an efficient decellularization method to generate. The whole procedure was carried out with special caution not to damage the glissons capsule, which surrounds the organ.
Keep the organ hydrated in phosphate buffered saline pbs in a 10cm petri dish. Heterotopic transplantation of a decellularized and. We will be able to use the organs of pigs surprisingly their organs have a high degree. By using a process called whole organ decellularization, scientists grew functioning heart tissue by. A recent innovation in tissue and organ engineering is the technique of whole organ decellularization, which allows the production of complex threedimensional extracellular matrix ecm bioscaffolds of the entire organ with preservation of the intrinsic vascular network. Developing a rapid, controllable, automated system for production of reproducible matrices in a closed system is a major step forward in whole organ tissue engineering. Decellularization and recellularization of whole livers. Scientists develop technique ensuring 100% success rate of. Recent studies have shown that decellularization of whole organs such as livers 4, 5, 28, 29, lung 19, 21, and heart are possible in rodent models. Carruthers, christopher arthur 2014 tissue and whole organ decellularization. The apparatus removes native cellular material from natural vascularized organs mechanically and chemically by perfusing detergents and wash buffers through an intact artery. Heterotopic transplantation of a decellularized and recellularized whole porcine heart. An overview of decellularization methods, their effect upon resulting ecm structure and composition, and recently described perfusion techniques for whole organ decellularization techniques are. In recent years, several studies have demonstrated the potential application of decellularization and recellularization approach for the development of functional bioartificial organs.
Organ and tissue transplantation treat a variety of medical problems, ranging from end. Comparable decellularization of fetal and adult cardiac. Decellularization and antibody staining of mouse tissues. The use of whole organ decellularization for the generation of a vascularized liver organoid pedro m. Hepatic tissue engineering using decellularized scaffolds is a potential. Can this organ transplant technique be humanitys hope to gaining immortality. We recently developed an ecmspecialized method termed in situ decellularization of tissues isdot to isolate native 3d ecm scaffolds from whole organs in which ecm structure and composition are. The vascular network remains intact even after full decellularization has been achieved, hence offering a route for the efficient delivery and penetration of cells and nutrients into the ecm scaffold during the process of recellularization to create a functional organ construct see fig. Comparison of methods for wholeorgan decellularization in.
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