Rapid Protocol of Porcine Kidney Decellularization

2018 ◽  
Vol 38 ◽  
pp. 67-74
Author(s):  
Fernanda Rocha de Souza ◽  
Maria Aparecida Dalboni ◽  
Andreas Kaasi ◽  
José Osmar Medina de Abreu Pestana ◽  
Adalberto Ramón Vieyra ◽  
...  
Keyword(s):  
Chronic Kidney Disease ◽  
Kidney Transplantation ◽  
Kidney Disease ◽  
Three Dimensional ◽  
Cell Types ◽  
Dimensional Structure ◽  
Porcine Kidney ◽  
Type Iv ◽  
Number Of Patients

Chronic kidney disease is a problem that has grown in recent decades worldwide. The National Kidney Foundation (NKF) estimates that the number of patients will double in the next 10 years. Dialysis and kidney transplantation are the treatments used for chronic kidney disease. There is hope in slowing down chronic kidney disease or even stopping its progression. Bioengineering and cell therapy are the main fields in kidney regeneration research using three-dimensional matrices in which cells are cultured, an ideal solution for scarcity organs for kidney transplantation. The difficulty in re-creating a functional kidney due to the complexity of its three-dimensional structure and its composition of different cell types and that can be incorporated in vivo with low immunogenicity is a very difficult task. Therefore, the aim of the present study was to meet the enormous demand for new treatments, developing strategies of tissue engineering on the basis of the decellularization of the porcine kidney performed through a new cell removal protocol. We determined the effective removal of cells by histologic and immunohistochemical analyses, showing the preservation of type IV collagen and fibronectin. Therefore, this method is a quick way to obtain decellularized porcine kidneys for future recellularization studies.

scholarly journals Dynamic reorganization of nuclear architecture during human cardiogenesis

2017 ◽  
Author(s):  
Paul A. Fields ◽  
Vijay Ramani ◽  
Giancarlo Bonora ◽  
Gurkan Yardimci ◽  
Alessandro Bertero ◽  
...  
Keyword(s):  
Pluripotent Stem Cells ◽  
Three Dimensional ◽  
Nuclear Architecture ◽  
Cell Types ◽  
Cardiac Differentiation ◽  
Dimensional Structure ◽  
Rna Seq ◽  
Dynamic Nature ◽  
Chromosomal Architecture

AbstractWhile chromosomal architecture varies among cell types, little is known about how this organization is established or its role in development. We integrated Hi-C, RNA-seq and ATAC-seq during cardiac differentiation from human pluripotent stem cells to generate a comprehensive profile of chromosomal architecture. We identified active and repressive domains that are dynamic during cardiogenesis and recapitulate in vivo cardiomyocytes. During differentiation, heterochromatic regions condense in cis. In contrast, many cardiac-specific genes, such as TTN (titin), decompact and transition to an active compartment coincident with upregulation. Moreover, we identify a network of genes, including TTN, that share the heart-specific splicing factor, RBM20, and become associated in trans during differentiation, suggesting the existence of a 3D nuclear splicing factory. Our results demonstrate both the dynamic nature in nuclear architecture and provide insights into how developmental genes are coordinately regulated.One Sentence SummaryThe three-dimensional structure of the human genome is dynamically regulated both globally and locally during cardiogenesis.

scholarly journals Interleukin-1α (IL-1α) is a Central Regulator of Leukocyte-Endothelial Adhesion in Myocardial Infarction and in Chronic Kidney Disease

Circulation ◽  
2021 ◽  
Author(s):  
Stefan J. Schunk ◽  
Sarah Triem ◽  
David Schmit ◽  
Stephen Zewinger ◽  
Tamim Sarakpi ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Chronic Kidney Disease ◽  
Endothelial Cells ◽  
Kidney Disease ◽  
Cell Types ◽  
Surface Expression ◽  
Organ Injury ◽  
Underlying Mechanisms

Background: Cardiovascular diseases (CVD) and chronic kidney disease (CKD) are highly prevalent, aggravate each other, and account for substantial mortality. Both conditions are characterized by activation of the innate immune system. The alarmin IL-1α is expressed in a variety of cell types promoting (sterile) systemic inflammation. The aim of the present study was to examine the role of IL-1α in mediating inflammation in the setting of acute myocardial infarction (AMI) and CKD. Methods: We assessed the expression of IL-1α on the surface of monocytes from patients with AMI and patients with CKD and determined its association with atherosclerotic CVD events during follow-up in an explorative clinical study. Furthermore, we assessed the inflammatory effects of IL-1α in several organ injury models in Il1a -/- and Il1b -/- mice and investigated the underlying mechanisms in vitro in monocytes and endothelial cells. Results: IL-1α is strongly expressed on the surface of monocytes from patients with AMI and CKD compared to healthy controls. Higher IL-1α surface expression on monocytes from patients with AMI and CKD was associated with a higher risk for atherosclerotic CVD events, which underlines the clinical relevance of IL-1α. In mice, IL-1α, but not IL-1β, mediates leukocyte-endothelial adhesion as determined by intravital microscopy. IL-1α promotes accumulation of macrophages and neutrophils in inflamed tissue in vivo . Furthermore, IL-1α on monocytes stimulates their homing at sites of vascular injury. A variety of stimuli such as free fatty acids or oxalate crystals induce IL-1α surface expression and release by monocytes, which then mediates their adhesion to the endothelium via IL-1 receptor-1. Besides, IL-1α promotes expression of the vascular cell adhesion molecule-1 (VCAM-1) on endothelial cells thereby fostering the adhesion of circulating leukocytes. IL-1α induces inflammatory injury after experimental AMI and abrogation of IL-1α prevents the development of CKD in oxalate or adenine-fed mice. Conclusions: IL-1α represents a key mediator of leukocyte-endothelial adhesion and inflammation in AMI and CKD. Inhibition of IL-1α may serve as a novel anti-inflammatory treatment strategy.

scholarly journals Adipokines as a link between obesity and chronic kidney disease

2013 ◽  
Vol 305 (12) ◽  
pp. F1629-F1636 ◽  
Author(s):  
Jessica F. Briffa ◽  
Andrew J. McAinch ◽  
Philip Poronnik ◽  
Deanne H. Hryciw
Keyword(s):  
Chronic Kidney Disease ◽  
Glomerular Filtration Rate ◽  
Kidney Disease ◽  
Glomerular Filtration ◽  
Filtration Rate ◽  
Cell Types ◽  
Peripheral Tissues ◽  
Increased Risk

Adipocytes secrete a number of bioactive adipokines that activate a variety of cell signaling pathways in central and peripheral tissues. Obesity is associated with the altered production of many adipokines and is linked to a number of pathologies. As an increase in body weight is directly associated with an increased risk for developing chronic kidney disease (CKD), there is significant interest in the link between obesity and renal dysfunction. Altered levels of the adipokines leptin, adiponectin, resistin, and visfatin can decrease the glomerular filtration rate and increase albuminuria, which are pathophysiological changes typical of CKD. Specifically, exposure of the glomerulus to altered adipokine levels can increase its permeability, fuse the podocytes, and cause mesangial cell hypertrophy, all of which alter the glomerular filtration rate. In addition, the adipokines leptin and adiponectin can act on tubular networks. Thus, adipokines can act on multiple cell types in the development of renal pathophysiology. Importantly, most studies have been performed using in vitro models, with future studies in vivo required to further elucidate the specific roles that adipokines play in the development and progression of CKD.

scholarly journals Dual Kidney Transplantation: A Review of Past and Prospect for Future

2017 ◽  
Vol 2017 ◽  
pp. 1-14 ◽  
Author(s):  
Muhammad Abdul Mabood Khalil ◽  
Jackson Tan ◽  
Taqi F. Toufeeq Khan ◽  
Muhammad Ashhad Ullah Khalil ◽  
Rabeea Azmat
Keyword(s):  
Chronic Kidney Disease ◽  
Kidney Transplantation ◽  
Kidney Disease ◽  
Treatment Options ◽  
Review Article ◽  
Donor Pool ◽  
Number Of Patients ◽  
History Of ◽  
Single Kidney ◽  
Day By Day

Kidney transplantation (KT) is one of the treatment options for patients with chronic kidney disease. The number of patients waiting for kidney transplantation is growing day by day. Various strategies have been put in place to expand the donor pool. Extended criteria donors are now accepted more frequently. Increasing number of elderly donors with age > 60 years, history of diabetes or hypertension, and clinical proteinuria are accepted as donor. Dual kidney transplantation (DKT) is also more frequently done and experience with this technique is slowly building up. DKT not only helps to reduce the number of patients on waiting list but also limits unnecessary discard of viable organs. Surgical complications of DKT are comparable to single kidney transplantation (SKT). Patient and graft survivals are also promising. This review article provides a summary of evidence available in the literature.

Epithelial Organotypic Cultures: A Viable Model to Address Mechanisms of Carcinogenesis by Epitheliotropic Viruses

2018 ◽  
Vol 18 (4) ◽  
pp. 246-255 ◽  
Author(s):  
Lara Termini ◽  
Enrique Boccardo
Keyword(s):  
Three Dimensional ◽  
Cell Types ◽  
Experimental Models ◽  
Biological Research ◽  
Specific Gene ◽  
Specific Cell ◽  
Organotypic Cultures ◽  
Skin Physiology

In vitro culture of primary or established cell lines is one of the leading techniques in many areas of basic biological research. The use of pure or highly enriched cultures of specific cell types obtained from different tissues and genetics backgrounds has greatly contributed to our current understanding of normal and pathological cellular processes. Cells in culture are easily propagated generating an almost endless source of material for experimentation. Besides, they can be manipulated to achieve gene silencing, gene overexpression and genome editing turning possible the dissection of specific gene functions and signaling pathways. However, monolayer and suspension cultures of cells do not reproduce the cell type diversity, cell-cell contacts, cell-matrix interactions and differentiation pathways typical of the three-dimensional environment of tissues and organs from where they were originated. Therefore, different experimental animal models have been developed and applied to address these and other complex issues in vivo. However, these systems are costly and time consuming. Most importantly the use of animals in scientific research poses moral and ethical concerns facing a steadily increasing opposition from different sectors of the society. Therefore, there is an urgent need for the development of alternative in vitro experimental models that accurately reproduce the events observed in vivo to reduce the use of animals. Organotypic cultures combine the flexibility of traditional culture systems with the possibility of culturing different cell types in a 3D environment that reproduces both the structure and the physiology of the parental organ. Here we present a summarized description of the use of epithelial organotypic for the study of skin physiology, human papillomavirus biology and associated tumorigenesis.

Kidney transplantation fails to provide adequate growth in children with chronic kidney disease born small for gestational age

2016 ◽  
Vol 32 (3) ◽  
pp. 511-519 ◽  
Author(s):  
Doris Franke ◽  
Rena Steffens ◽  
Lena Thomas ◽  
Leo Pavičić ◽  
Thurid Ahlenstiel ◽  
...  
Keyword(s):  
Chronic Kidney Disease ◽  
Kidney Transplantation ◽  
Kidney Disease ◽  
Gestational Age ◽  
Small For Gestational Age

scholarly journals Structure of the Lectin Mannose 6-Phosphate Receptor Homology (MRH) Domain of Glucosidase II, an Enzyme That Regulates Glycoprotein Folding Quality Control in the Endoplasmic Reticulum

2013 ◽  
Vol 288 (23) ◽  
pp. 16460-16475 ◽  
Author(s):  
Linda J. Olson ◽  
Ramiro Orsi ◽  
Solana G. Alculumbre ◽  
Francis C. Peterson ◽  
Ivan D. Stigliano ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Three Dimensional ◽  
Binding Pocket ◽  
Dimensional Structure ◽  
Mannose Residue ◽  
Glucosidase Ii ◽  
Mannose 6 Phosphate ◽  
First Time ◽  
Conserved Residue

Here we report for the first time the three-dimensional structure of a mannose 6-phosphate receptor homology (MRH) domain present in a protein with enzymatic activity, glucosidase II (GII). GII is involved in glycoprotein folding in the endoplasmic reticulum. GII removes the two innermost glucose residues from the Glc3Man9GlcNAc2 transferred to nascent proteins and the glucose added by UDP-Glc:glycoprotein glucosyltransferase. GII is composed of a catalytic GIIα subunit and a regulatory GIIβ subunit. GIIβ participates in the endoplasmic reticulum localization of GIIα and mediates in vivo enhancement of N-glycan trimming by GII through its C-terminal MRH domain. We determined the structure of a functional GIIβ MRH domain by NMR spectroscopy. It adopts a β-barrel fold similar to that of other MRH domains, but its binding pocket is the most shallow known to date as it accommodates a single mannose residue. In addition, we identified a conserved residue outside the binding pocket (Trp-409) present in GIIβ but not in other MRHs that influences GII glucose trimming activity.

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