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Perfusion decellularization

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https://www.readbyqxmd.com/read/29127860/human-embryonic-stem-cell-derived-cardiovascular-progenitor-cells-efficiently-colonize-in-bfgf-tethered-natural-matrix-to-construct-contracting-humanized-rat-hearts
#1
Sarah Rajabi, Sara Pahlavan, Mohammad Kazemi Ashtiani, Hassan Ansari, Saeed Abbasalizadeh, Forough Azam Sayahpour, Fahimeh Varzideh, Sawa Kostin, Nasser Aghdami, Thomas Braun, Hossein Baharvand
Bioengineering of whole hearts using human embryonic stem cells (hESCs)-derived cardiovascular progenitor cells (CPCs) and natural matrices is a promising approach to overcome organ donor shortage threatening millions of patients awaiting for heart transplantation. Here, we developed a novel strategy for generation of heart constructs by repopulating engineered decellularized rat hearts using hESCs-derived CPCs. Careful expansion of CPCs in a scalable stirred-suspension bioreactor combined with step-wise seeding (60 million cells in 3 steps of 20 million per 1...
November 1, 2017: Biomaterials
https://www.readbyqxmd.com/read/29118967/cold-perfusion-decellularization-of-whole-organ-porcine-pancreas-supports-human-fetal-pancreatic-cell-attachment-and-expression-of-endocrine-and-exocrine-markers
#2
Erik Elebring, Vijay K Kuna, Niclas Kvarnström, Suchitra Sumitran-Holgersson
Despite progress in the field of decellularization and recellularization, the outcome for pancreas has not been adequate. This might be due to the challenging dual nature of pancreas with both endocrine and exocrine tissues. We aimed to develop a novel and efficient cold-perfusion method for decellularization of porcine pancreas and recellularize acellular scaffolds with human fetal pancreatic stem cells. Decellularization of whole porcine pancreas at 4°C with sodium deoxycholate, Triton X-100 and DNase efficiently removed cellular material, while preserving the extracellular matrix structure...
January 2017: Journal of Tissue Engineering
https://www.readbyqxmd.com/read/29102342/bioengineered-lungs-a-challenge-and-an-opportunity
#3
Ramon Farré, Jordi Otero, Isaac Almendros, Daniel Navajas
Lung biofabrication is a new tissue engineering and regenerative development aimed at providing organs for potential use in transplantation. Lung biofabrication is based on seeding cells into an acellular organ scaffold and on culturing them in an especial purpose bioreactor. The acellular lung scaffold is obtained by decellularizing a non-transplantable donor lung by means of conventional procedures based on application of physical, enzymatic and detergent agents. To avoid immune recipient's rejection of the transplanted bioengineered lung, autologous bone marrow/adipose tissue-derived mesenchymal stem cells, lung progenitor cells or induced pluripotent stem cells are used for biofabricating the bioengineered lung...
November 1, 2017: Archivos de Bronconeumología
https://www.readbyqxmd.com/read/29101679/simple-and-quick-method-to-obtain-a-decellularized-functional-liver-bioscaffold
#4
Matteo Ghiringhelli, Alessandro Zenobi, Stefano Brizzola, Fulvio Gandolfi, Valentino Bontempo, Sandro Rossi, Tiziana A L Brevini, Fabio Acocella
The development of new approaches for organ transplantation has become crucial in the last years. In particular, organ engineering, involving the preparation of acellular matrices that provide a natural habitat for reseeding with an appropriate population of cells, is an attractive although technically demanding approach. We here describe a method that allows for the derivation of functional in vitro hepatic organoids and that does not require a previous selection of all the parenchymal hepatocytes and non-parenchymal cells, namely, Kupffer cells, liver endothelial cells, and hepatic stellate cells...
November 4, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29101678/liver-bioengineering-using-decellularized-whole-liver-scaffolds
#5
Iris Pla-Palacín, Pilar Sainz-Arnal, Sara Morini, Manuel Almeida, Pedro M Baptista
Currently, due to the progress made in the field of regenerative medicine, whole-organ bioengineering is becoming a valid alternative to cope with the shortages of organs for transplantation. In this chapter, we describe the main techniques carried out for pig liver bioengineering, which serves as an essential model for future human liver bioengineering. These include porcine whole-liver decellularization, endothelial and mesenchymal stem cell isolation, porcine ES-derived hepatoblasts, and scaffold recellularization using a bioreactor perfusion system...
November 4, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29091040/a-pilot-study-to-evaluate-the-effects-of-perfusion-decellularized-porcine-hepatic-derived-wound-matrix-on-difficult-to-heal-diabetic-foot-ulcers
#6
Robert Fridman, Jonathan Engelhardt
OBJECTIVE: The purpose of this prospective pilot study is to evaluate the effects of a novel perfusion-decellularized porcine hepatic-derived wound matrix (PDPHD-WM) on very difficult-to-heal diabetic foot ulcers (DFUs). MATERIALS AND METHODS: For the intent of this study, a very difficult-to-heal DFU is one with a duration > 3 months despite using at least 1 advanced healing modality. Seven patients were enrolled in the study for a length of 14 weeks, including a 2-week run-in phase of standard-of-care therapy with saline gel and offloading in a CAM walker...
October 2017: Wounds: a Compendium of Clinical Research and Practice
https://www.readbyqxmd.com/read/29081492/tissue-engineered-small-vessel-conduits-the-anti-thrombotic-effect-of-re-endothelialization-of-decellularized-baboon-arteries-a-preliminary-experimental-study
#7
Muriel Meiring, Mmakgabu Khemisi, Leana Laker, Pascal M Dohmen, Francis E Smit
BACKGROUND The use of decellularized biological scaffolds for the reconstruction of small-diameter vascular grafts remains a challenge in tissue engineering. Thrombogenicity is an important cause of obstruction in these vessels due to decellularization. Seeding of the decellularized vascular constructs with endothelial cells is therefore a prerequisite for the prevention of thrombosis. The aim of this study was to seed decellularized baboon arteries with endothelial cells and to compare the thrombogenicity to that of decellularized arteries after circulation of blood...
October 30, 2017: Medical Science Monitor Basic Research
https://www.readbyqxmd.com/read/29079859/evaluation-of-decellularized-porcine-jejunum-as-a-matrix-for-lacrimal-gland-reconstruction-in-vitro-for-treatment-of-dry-eye-syndrome
#8
Isobel Massie, Kristina Spaniol, Andreas Barbian, Gereon Poschmann, Kai Stühler, Gerd Geerling, Marco Metzger, Sonja Mertsch, Stefan Schrader
Purpose: Dry eye syndrome (DES) can cause blindness in severe cases, but mainly palliative treatments exist. A tissue-engineered lacrimal gland (LG) could provide a curative treatment. We aimed to evaluate decellularized porcine jejunum (SIS-Muc) as a scaffold for porcine LG epithelial cells. Methods: To evaluate SIS-Muc as a potential scaffold, basement membrane proteins in SIS-Muc and native LG were compared (immunohistochemistry [IHC]). Porcine LG epithelial cells cultured on plastic were characterized (immunocytochemistry), and their culture supernatant was compared with porcine tears (proteomics)...
October 1, 2017: Investigative Ophthalmology & Visual Science
https://www.readbyqxmd.com/read/29018244/bioengineering-of-functional-human-induced-pluripotent-stem-cell-derived-intestinal-grafts
#9
Kentaro Kitano, Dana M Schwartz, Haiyang Zhou, Sarah E Gilpin, Gregory R Wojtkiewicz, Xi Ren, Cesar A Sommer, Amalia V Capilla, Douglas J Mathisen, Allan M Goldstein, Gustavo Mostoslavsky, Harald C Ott
Patients with short bowel syndrome lack sufficient functional intestine to sustain themselves with enteral intake alone. Transplantable vascularized bioengineered intestine could restore nutrient absorption. Here we report the engineering of humanized intestinal grafts by repopulating decellularized rat intestinal matrix with human induced pluripotent stem cell-derived intestinal epithelium and human endothelium. After 28 days of in vitro culture, hiPSC-derived progenitor cells differentiate into a monolayer of polarized intestinal epithelium...
October 10, 2017: Nature Communications
https://www.readbyqxmd.com/read/29017397/liver-scaffolds-support-survival-and-metabolic-function-of-multilineage-neonatal-allogenic-cells
#10
Wessam Hassanein, Arielle Cimeno, Avraham Werdesheim, Bryan Buckingham, Joshua Harrison, Mehmet C Uluer, Ali Khalifeh, Carlos Rivera-Pratts, Stephen Klepfer, Jhade D Woodall, Urmil Dhru, Elliot Bromberg, Dawn Parsell, Cinthia Drachenberg, Rolf N Barth, John C LaMattina
Organ scaffold bioengineering is currently limited by the inability to effectively repopulate the scaffold with appropriately distributed functional cells. We examined the feasibility of a decellularized liver scaffold to support the growth and function of multilineage allogenic cells derived from either adult or neonatal liver cells. Cell slurries from neonatal and adult rat livers containing hepatocytes, cholangiocytes, and endothelial cells were introduced into decellularized adult rat liver scaffolds via the bile duct...
October 10, 2017: Tissue Engineering. Part A
https://www.readbyqxmd.com/read/28922323/face-graft-scaffold-production-in-a-rat-model
#11
Jérôme Duisit, Hadrien Amiel, Giuseppe Orlando, Adeline Dedriche, Catherine Behets, Pierre Gianello, Benoît Lengelé
BACKGROUND: As a route towards face bioengineering, our group previously reported the production of a complete scaffold production by perfusion-decellularization of a porcine ear subunit graft and partial recellularization. In order to extend the scaffold to the whole face and to down-scale it, we applied our findings to a rodent hemi-facial graft model. METHODS: After euthanasia, seven full-thickness rat hemi-face grafts were harvested with the CCA and the EJV as pedicle, and cannulated...
September 14, 2017: Plastic and Reconstructive Surgery
https://www.readbyqxmd.com/read/28895502/characterization-of-the-epicardial-adipose-tissue-in-decellularized-human-scaled-whole-hearts-implications-for-the-whole-heart-tissue-engineering
#12
Payam Akhyari, Fabian Oberle, Joern Huelsmann, Hans Heid, Stefan Lehr, Andreas Barbian, Sentaro Nakanishi, Hug Aubin, Alexander Jenke, Artur Lichtenberg
Whole-organ engineering is an innovative field of regenerative medicine with growing translational perspectives. Recent reports suggest the feasibility of decellularization and repopulation of entire human size hearts. However, little is known about the susceptibility of epicardial adipose tissue (EAT) to decellularization. Here, human size hearts of ovine donors were subjected to perfusion-based decellularization using detergent solutions. Upon basic histological evaluation and total DNA measurement myocardial regions prove largely decellularized while EAT demonstrated cellular remnants, further confirmed by transmission electron microscopy...
September 12, 2017: Tissue Engineering. Part A
https://www.readbyqxmd.com/read/28886732/optimized-protocol-for-whole-organ-decellularization
#13
A Schmitt, R Csiki, A Tron, B Saldamli, J Tübel, K Florian, S Siebenlist, E Balmayor, R Burgkart
BACKGROUND: The idea of tissue decellularization to gain matrices for tissue engineering is promising. The aim of the present study is to establish a safe and reproducible protocol for solid tissue decellularization that prevents the architecture of the matrix with the inherent vascular network. METHODS: The study was performed in rat kidneys which were decellularized by a SDS-based perfusion protocol. Perfusion time and SDS concentration were systematically changed to obtain the shortest and most gentle protocol that leads to complete decellularization...
September 8, 2017: European Journal of Medical Research
https://www.readbyqxmd.com/read/28863041/decellularized-rat-lung-scaffolds-using-sodium-lauryl-ether-sulfate-for-tissue-engineering
#14
Jinhui Ma, Zhihai Ju, Jie Yu, Yeru Qiao, Chenwei Hou, Chen Wang, Feilong Hei
Perfusion decellularization with detergents is effective to maintain the architecture and proteins of extracellular matrix (ECM) for use in the field of lung tissue engineering (LTE). However, it is unclear which detergent is ideal to produce an acellular lung scaffold. In this study, we obtained two decellularized rat lung scaffolds using a novel detergent sodium lauryl ether sulfate (SLES) and a conventional detergent sodium dodecyl sulfate (SDS). Both decellularized lung scaffolds were assessed by histology, immunohistochemistry, scanning electron microscopy, DNA quantification, sulfated glycosaminoglycans (GAGs) quantification and western blot...
August 31, 2017: ASAIO Journal: a Peer-reviewed Journal of the American Society for Artificial Internal Organs
https://www.readbyqxmd.com/read/28828244/freeze-thaw-decellularization-of-the-trabecular-meshwork-in-an-ex-vivo-eye-perfusion-model
#15
Yalong Dang, Susannah Waxman, Chao Wang, Adrianna Jensen, Ralitsa T Loewen, Richard A Bilonick, Nils A Loewen
OBJECTIVE: The trabecular meshwork (TM) is the primary substrate of outflow resistance in glaucomatous eyes. Repopulating diseased TM with fresh, functional TM cells might be a viable therapeutic approach. Decellularized TM scaffolds have previously been produced by ablating cells with suicide gene therapy or saponin, which risks incomplete cell removal or dissolution of the extracellular matrix, respectively. We hypothesized that improved trabecular meshwork cell ablation would result from freeze-thaw cycles compared to chemical treatment...
2017: PeerJ
https://www.readbyqxmd.com/read/28776178/protocols-for-rat-uterus-isolation-and-decellularization-applications-for-uterus-tissue-engineering-and-3d-cell-culturing
#16
Arvind Manikantan Padma, Tom Tristan Tiemann, Ahmed Baker Alshaikh, Randa Akouri, Min Jong Song, Mats Hellström
Sophisticated culturing conditions are required to grow cells in a three-dimensional (3D) environment. Cells then require a type of scaffold rich in proteins, growth factors, and signaling molecules that simulates their natural environment. Tissues from all species of animals have an organ-specific extracellular matrix (ECM) structure that plays a key role in cell proliferation and migration. Hence, the scaffold composition plays a significant role for any successful 3D cell culturing system. We developed a whole rat uterus ECM scaffold by the perfusion of detergents and ionic solutions through the vascular system of an isolated normal rat uterus in a process termed "decellularization...
August 4, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28758007/re-epithelialization-of-whole-porcine-kidneys-with-renal-epithelial-cells
#17
Nafiseh Poornejad, Evan Buckmiller, Lara Schaumann, Haonan Wang, Jonathan Wisco, Beverly Roeder, Paul Reynolds, Alonzo Cook
Decellularized porcine kidneys were recellularized with renal epithelial cells by three methods: perfusion through the vasculature under high pressure, perfusion through the ureter under high pressure, or perfusion through the ureter under moderate vacuum. Histology, scanning electron microscopy, confocal microscopy, and magnetic resonance imaging were used to assess vasculature preservation and the distribution of cells throughout the kidneys. Cells were detected in the magnetic resonance imaging by labeling them with iron oxide...
January 2017: Journal of Tissue Engineering
https://www.readbyqxmd.com/read/28742686/bioengineering-a-human-face-graft-the-matrix-of-identity
#18
Jérôme Duisit, Louis Maistriaux, Adriano Taddeo, Giuseppe Orlando, Virginie Joris, Emmanuel Coche, Catherine Behets, Jan Lerut, Chantal Dessy, Giulio Cossu, Esther Vögelin, Robert Rieben, Pierre Gianello, Benoît Lengelé
OBJECTIVE: During the last decade, face allotransplantation has been shown to be a revolutionary reconstructive procedure for severe disfigurements. However, offer to patients remains limited due to lifelong immunosuppression. To move forward in the field, a new pathway in tissue engineering is proposed. BACKGROUND: Our previously reported technique of matrix production of a porcine auricular subunit graft has been translated to a human face model. METHODS: 5 partial and 1 total face grafts were procured from human fresh cadavers...
November 2017: Annals of Surgery
https://www.readbyqxmd.com/read/28735385/preparation-of-decellularized-liver-scaffolds-and-recellularized-liver-grafts
#19
Yibin Chen, Sharon Geerts, Maria Jaramillo, Basak E Uygun
Severe liver disease is the 12th leading cause of death in the USA, with organ transplantation often being the only viable option for treatment. However, due to the shortage of viable donor livers, it is estimated that over 1200 patients died in 2015 while waiting for liver transplantation. This highlights the need for alternative sources of viable organs. In this study, we describe a method that provides the groundwork for the development of functional liver grafts. The approach described here is for removal of cells from intact livers and subsequently repopulating them with functional liver cells...
July 23, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28726588/reconstituting-mouse-lungs-with-conditionally-reprogrammed-human-bronchial-epithelial-cells
#20
Ryan LaRanger, Jennifer R Peters-Hall, Melissa Coquelin, Busola R Alabi, Christopher T Chen, Woodring E Wright, Jerry W Shay
We developed methods for conditionally reprogramming (CR) primary human bronchial epithelial cells (HBECs) to extend their functional lifespan and permit their differentiation into both upper and lower airway lung epithelium. We also developed a bioreactor to support vascular perfusion and rhythmic breathing of decellularized mouse lungs reconstituted with CR HBECs isolated from patients with and without cystic fibrosis (CF). While conditionally reprogrammed cells only differentiate into an upper airway epithelium after 35 days at the air-liquid interface, in reconstituted lungs these cells differentiate into upper airway bronchial epithelium and lower airway alveolar structures after 12 days...
September 25, 2017: Tissue Engineering. Part A
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