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Perfusion decellularization

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https://www.readbyqxmd.com/read/28029762/prevascularization-of-decellularized-porcine-myocardial-slice-for-cardiac-tissue-engineering
#1
Pawan Kc, Mickey Shah, Jun Liao, Ge Zhang
Prevacularization strategies have been implemented in tissue engineering to generate microvasculature networks within a scaffold prior to implantation. Prevascularizing scaffolds will shorten the time of functional vascular perfusion with host upon implantation. In this study, we explored key variables affecting the interaction between decellularized porcine myocardium slices (dPMSs) and reseeded stem cells toward the fabrication of prevascularized cardiac tissue. Our results demonstrated that dPMS supports attachment of human mesenchymal stem cells (hMSCs) and rat adipose derived stem cells (rASCs) with high viability...
January 10, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28029279/recellularization-via-the-bile-duct-supports-functional-allogenic-and-xenogenic-cell-growth-on-a-decellularized-rat-liver-scaffold
#2
Wessam Hassanein, Mehmet C Uluer, John Langford, Jhade D Woodall, Arielle Cimeno, Urmil Dhru, Avraham Werdesheim, Joshua Harrison, Carlos Rivera-Pratt, Stephen Klepfer, Ali Khalifeh, Bryan Buckingham, Philip S Brazio, Dawn Parsell, Charlie Klassen, Cinthia Drachenberg, Rolf N Barth, John C LaMattina
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)...
December 28, 2016: Organogenesis
https://www.readbyqxmd.com/read/28019665/the-impact-of-left-ventricular-stretching-in-model-cultivations-with-neonatal-cardiomyocytes-in-a-whole-heart-bioreactor
#3
Jörn Hülsmann, Hug Aubin, Alexander Wehrmann, Artur Lichtenberg, Payam Akhyari
Here, we investigate the impact of integrated three-dimensional (3D) left ventricular (LV) stretching on myocardial maturation in a whole-heart bioreactor setting. Therefore, decellularized rat hearts were selectively repopulated with rodent neonatal cardiomyocytes (5•10(6) cells per heart) and cultured over 5 d. Continuous medium perfusion was maintained through the coronary artery system in a customized whole-heart bioreactor system with or without integrated biomechanical stimulation of LV. 3D repopulation effectiveness and cellular vitality were evaluated by repetitive metabolic WST-1 assays and 3D confocal microscopy analysis through fluorescent staining, also assessing cellular organization...
December 26, 2016: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/27957815/evolution-of-graft-morphology-and-function-after-recellularization-of-decellularized-rat-livers
#4
Antje Butter, Khalid Aliyev, Karl-Herbert Hillebrandt, Nathanael Raschzok, Martin Kluge, Nicolai Seiffert, Peter Tang, Hendrik Napierala, Muhammad Imtiaz Ashraf, Anja Reutzel-Selke, Andreas Andreou, Johann Pratschke, Igor Maximilian Sauer, Benjamin Struecker
Decellularization of livers is a well-established procedure. Data on different reseeding techniques or the functional evolution and re-organization processes of repopulated grafts remains limited. We established a proprietary, customized bioreactor to repopulate decellularized rat livers (n = 21) with primary rat hepatocytes (150 x 10(6) cells) via the hepatic artery and to subsequently evaluate graft morphology and function during seven days of ex vivo perfusion. Grafts were analyzed at 1 h, 6 h, 12 h, 24 h, 3d, 5d and 7d after recellularization (all n = 3) by immuno-histologic evaluation, hepatocyte-related enzyme (AST, ALT, LDH) and albumin measurement in the perfusate...
December 13, 2016: Journal of Tissue Engineering and Regenerative Medicine
https://www.readbyqxmd.com/read/27943597/re-endothelialization-of-rat-lung-scaffolds-through-passive-gravity-driven-seeding-of-segment-specific-pulmonary-endothelial-cells
#5
Michelle E Scarritt, Nicholas C Pashos, Jessica M Motherwell, Zachary R Eagle, Brian J Burkett, Ashley N Gregory, Ricardo Mostany, Daniel J Weiss, Diego F Alvarez, Bruce A Bunnell
Effective re-endothelialization is critical for the use of decellularized scaffolds for ex vivo lung engineering. Current approaches yield insufficiently re-endothelialized scaffolds that hemorrhage and become thrombogenic upon implantation. Herein, gravity-driven seeding coupled with bioreactor culture facilitated widespread distribution and engraftment of endothelial cells throughout rat lung scaffolds. Initially, human umbilical vein endothelial cells (HUVECs) were seeded into the pulmonary artery by either gravity-driven, variable flow perfusion seeding or pump-driven, pulsatile flow perfusion seeding...
December 12, 2016: Journal of Tissue Engineering and Regenerative Medicine
https://www.readbyqxmd.com/read/27942003/biochemical-and-cellular-determinants-of-renal-glomerular-elasticity
#6
Addie E Embry, Hamid Mohammadi, Xinying Niu, Liping Liu, Borren Moe, William A Miller-Little, Christopher Y Lu, Leslie A Bruggeman, Christopher A McCulloch, Paul A Janmey, R Tyler Miller
The elastic properties of renal glomeruli and their capillaries permit them to maintain structural integrity in the presence of variable hemodynamic forces. Measured by micro-indentation, glomeruli have an elastic modulus (E, Young's modulus) of 2.1 kPa, and estimates from glomerular perfusion studies suggest that the E of glomeruli is between 2 and 4 kPa. F-actin depolymerization by latrunculin, inhibition of acto-myosin contractility by blebbistatin, reduction in ATP synthesis, and reduction of the affinity of adhesion proteins by EDTA reduced the glomerular E to 1...
2016: PloS One
https://www.readbyqxmd.com/read/27924607/fabrication-and-characterization-of-decellularized-periodontal-ligament-cell-sheet-constructs
#7
Amro Farag, Cédryck Vaquette, Dietmar W Hutmacher, P Mark Bartold, Saso Ivanovski
Decellularized tissue-engineered constructs have the potential to promote regeneration by providing a biomimetic extracellular matrix that directs tissue-specific regeneration when implanted in situ. Recently, the use of cell sheets has shown promising results in promoting periodontal regeneration. Here, we describe the fabrication of decellularized periodontal cell sheets with intact extracellular matrix structural and biological properties. Melt electro spun polycaprolactone (PCL) scaffolds are used as a carrier for the inherently fragile cell sheets, to provide support during the processes of decellularization...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27911373/the-mouse-isolated-perfused-kidney-technique
#8
Jan Czogalla, Frank Schweda, Johannes Loffing
The mouse isolated perfused kidney (MIPK) is a technique for keeping a mouse kidney under ex vivo conditions perfused and functional for 1 hr. This is a prerequisite for studying the physiology of the isolated organ and for many innovative applications that may be possible in the future, including perfusion decellularization for kidney bioengineering or the administration of anti-rejection or genome-editing drugs in high doses to prime the kidney for transplantation. During the time of the perfusion, the kidney can be manipulated, renal function can be assessed, and various pharmaceuticals administered...
November 17, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27911030/acellular-endocardium-as-a-novel-biomaterial-for-the-intima-of-tissue-engineered-small-caliber-vascular-grafts
#9
Feng Wang, Xin Guan, TianYi Wu, JianOu Qiao, ZhaoQing Han, JinLong Wu, XiaoWei Yu, QingJun You
We aimed to investigate whether acellular endocardium can be used as a useful biomaterial for the intima of engineered small-caliber vascular grafts. Fresh endocardium was harvested from the swine left atrium and was decellularized by digestion with the decellularization solution of Triton X-100 and SDS containing DNase I and RNase A. Surface morphological characteristics and Young's modulus were evaluated. To analyze the effect of mechanical characteristics on cell adhesion, the decellularized endocardium was stiffened with 2...
December 2016: Artificial Organs
https://www.readbyqxmd.com/read/27894293/lung-bioengineering-physical-stimuli-and-stem-progenitor-cell-biology-interplay-towards-biofabricating-a-functional-organ
#10
REVIEW
Paula N Nonaka, Juan J Uriarte, Noelia Campillo, Vinicius R Oliveira, Daniel Navajas, Ramon Farré
A current approach to obtain bioengineered lungs as a future alternative for transplantation is based on seeding stem cells on decellularized lung scaffolds. A fundamental question to be solved in this approach is how to drive stem cell differentiation onto the different lung cell phenotypes. Whereas the use of soluble factors as agents to modulate the fate of stem cells was established from an early stage of the research with this type of cells, it took longer to recognize that the physical microenvironment locally sensed by stem cells (e...
November 28, 2016: Respiratory Research
https://www.readbyqxmd.com/read/27884776/inverted-orientation-improves-decellularization-of-whole-porcine-hearts
#11
Po-Feng Lee, Eric Chau, Rafael Cabello, Alvin T Yeh, Luiz C Sampaio, Andrea S Gobin, Doris A Taylor
: In structurally heterogeneous organs, such as heart, it is challenging to retain extracellular matrix integrity in the thinnest regions (eg, valves) during perfusion decellularization and completely remove cellular debris from thicker areas. The high inflow rates necessary to maintain physiologic pressure can distend or damage thin tissues, but lower pressures prolong the process and increase the likelihood of contamination. We examined two novel retrograde decellularization methods for porcine hearts: inverting the heart or venting the apex to decrease inflow rate...
November 21, 2016: Acta Biomaterialia
https://www.readbyqxmd.com/read/27871686/eliminating-the-organ-transplant-waiting-list-the-future-with-perfusion-decellularized-organs
#12
REVIEW
Dominique Seetapun, Jeffrey J Ross
No abstract text is available yet for this article.
November 18, 2016: Surgery
https://www.readbyqxmd.com/read/27862079/discarded-livers-find-a-new-life-engineered-liver-grafts-using-hepatocytes-recovered-from-marginal-livers
#13
Basak E Uygun, Maria-Louisa Izamis, Maria Jaramillo, Yibin Chen, Gavrielle Price, Sinan Ozer, Martin L Yarmush
Treatment for end-stage liver failure is restricted by the critical shortage of donor organs; about 4000 people die in the USA while waiting for a transplantable organ. This situation has been a major driving force behind the rise of tissue engineering to build artificial tissues/organs. Recent advancements in creating transplantable liver grafts using decellularized liver scaffolds bring the field closer to clinical translation. However, a source of readily available and highly functional adult hepatocytes in adequate numbers for regenerative liver therapies still remains unclear...
November 8, 2016: Artificial Organs
https://www.readbyqxmd.com/read/27826794/the-challenge-in-using-mesenchymal-stromal-cells-for-recellularization-of-decellularized-cartilage
#14
REVIEW
Zhao Huang, Owen Godkin, Gundula Schulze-Tanzil
Some decellularized musculoskeletal extracellular matrices (ECM)s derived from tissues such as bone, tendon and fibrocartilaginous meniscus have already been clinical use for tissue reconstruction. Repair of articular cartilage with its unique zonal ECM architecture and composition is still an unsolved problem, and the question is whether allogenic or xenogeneic decellularized cartilage ECM could serve as a biomimetic scaffold for this purpose.Hence, this survey outlines the present state of preparing decellularized cartilage ECM-derived scaffolds or composites for reconstruction of different cartilage types and of reseeding it particularly with mesenchymal stromal cells (MSCs)...
November 8, 2016: Stem Cell Reviews
https://www.readbyqxmd.com/read/27805597/study-of-viral-vectors-in-a-three-dimensional-liver-model-repopulated-with-the-human-hepatocellular-carcinoma-cell-line-hepg2
#15
Thomas Hiller, Viola Röhrs, Eva-Maria Dehne, Anke Wagner, Henry Fechner, Roland Lauster, Jens Kurreck
This protocol describes the generation of a three-dimensional (3D) ex vivo liver model and its application to the study and development of viral vector systems. The model is obtained by repopulating the extracellular matrix of a decellularized rat liver with a human hepatocyte cell line. The model permits studies in a vascularized 3D cell system, replacing potentially harmful experiments with living animals. Another advantage is the humanized nature of the model, which is closer to human physiology than animal models...
October 24, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27784199/dynamic-autologous-reendothelialization-of-small-caliber-arterial-extracellular-matrix-a-preclinical-large-animal-study
#16
Nitsan Dahan, Udi Sarig, Tomer Bronshtein, Limor Baruch, Tony Karram, Aaron Hoffman, Marcelle Machluf
Effective cellularization is a key approach to prevent small-caliber (<4 mm) tissue-engineered vascular graft (TEVG) failure and maintain patency and contractility following implantation. To achieve this goal, however, improved biomimicking designs and/or relatively long production times (typically several months) are required. We previously reported on porcine carotid artery decellularization yielding biomechanically stable and cell supportive small-caliber (3-4 mm diameter, 5 cm long) arterial extracellular matrix (scaECM) vascular grafts...
January 2017: Tissue Engineering. Part A
https://www.readbyqxmd.com/read/27720934/in-vivo-organ-engineering-perfusion-of-hepatocytes-in-a-single-liver-lobe-scaffold-of-living-rats
#17
Jun Pan, Sheng Yan, Jun-Jie Gao, Ying-Ying Wang, Zhong-Jie Lu, Chen-Wei Cui, Yao-Hui Zhang, Yan Wang, Xue-Qin Meng, Lin Zhou, Hai-Yang Xie, Jessica Zheng, Ming H Zheng, Shu-Sen Zheng
Organ decellularization is emerging as a promising regenerative medicine approach as it is able to provide an acellular, three-dimensional biological scaffold material that can be seeded with living cells for organ reengineering. However this application is currently limited to donor-derived decellularized organs for reengineering in vitro and no study has been conducted for re-engineering the decellularized organ in vivo. We developed a novel technique of a single liver lobe decellularization in vivo in live animals...
October 5, 2016: International Journal of Biochemistry & Cell Biology
https://www.readbyqxmd.com/read/27676406/investigating-neovascularization-in-rat-decellularized-intestine-an-in-vitro-platform-for-studying-angiogenesis
#18
Lindsey Dew, William R English, Chuh K Chong, Sheila MacNeil
One of the main challenges currently faced by tissue engineers is the loss of tissues postimplantation due to delayed neovascularization. Several strategies are under investigation to create vascularized tissue, but none have yet overcome this problem. In this study, we produced a decellularized natural vascular scaffold from rat intestine to use as an in vitro platform for neovascularization studies for tissue-engineered constructs. Decellularization resulted in almost complete (97%) removal of nuclei and DNA, while collagen, glycosaminoglycan, and laminin content were preserved...
December 2016: Tissue Engineering. Part A
https://www.readbyqxmd.com/read/27622532/regenerative-potential-of-human-airway-stem-cells-in-lung-epithelial-engineering
#19
Sarah E Gilpin, Jonathan M Charest, Xi Ren, Luis F Tapias, Tong Wu, Daniele Evangelista-Leite, Douglas J Mathisen, Harald C Ott
Bio-engineered organs for transplantation may ultimately provide a personalized solution for end-stage organ failure, without the risk of rejection. Building upon the process of whole organ perfusion decellularization, we aimed to develop novel, translational methods for the recellularization and regeneration of transplantable lung constructs. We first isolated a proliferative KRT5(+)TP63(+) basal epithelial stem cell population from human lung tissue and demonstrated expansion capacity in conventional 2D culture...
November 2016: Biomaterials
https://www.readbyqxmd.com/read/27459202/bioengineered-livers-a-new-tool-for-drug-testing-and-a-promising-solution-to-meet-the-growing-demand-for-donor-organs
#20
Franziska Mußbach, Utz Settmacher, Olaf Dirsch, Chichi Xie, Uta Dahmen
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...
2016: European Surgical Research. Europäische Chirurgische Forschung. Recherches Chirurgicales Européennes
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