journal
MENU ▼
Read by QxMD icon Read
search

Methods in Molecular Biology

journal
https://www.readbyqxmd.com/read/28735385/preparation-of-decellularized-liver-scaffolds-and-recellularized-liver-grafts
#1
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/28730534/tracheal-cartilage-isolation-and-decellularization
#2
Zachary Galliger, Angela Panoskaltsis-Mortari
Tissue decellularization allows isolation of tissue extracellular matrix components, enabling bioengineering of native tissue microenvironments with minimal antigenic components. Here we describe a method to harvest decellularized cartilage tissue from donor trachea using a series of chemical and enzymatic washes and incorporating the extracellular matrix in gelatin methacrylate hydrogels. This decellularized cartilage tissue is easily incorporated into a variety of hydrogels to create a cartilage tissue scaffold...
July 21, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28710687/disaggregation-and-reaggregation-of-zebrafish-retinal-cells-for-the-analysis-of-neuronal-layering
#3
Megan K Eldred, Leila Muresan, William A Harris
The reaggregation of dissociated cells to form organotypic structures provides an in vitro system for the analysis of the cellular interactions and molecular mechanisms involved in the formation of tissue architecture. The retina, an outgrowth of the forebrain, is a precisely layered neural tissue, yet the mechanisms underlying layer formation are largely unexplored. Here we describe the protocol to dissociate, re-aggregate, and culture zebrafish retinal cells from a transgenic, Spectrum of Fates, line where all main cell types are labelled with a combination of fluorescent proteins driven by fate-specific promoters...
July 15, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28702885/use-of-a-super-hydrophobic-microbioreactor-to-generate-and-boost-pancreatic-mini-organoids
#4
Tiziana A L Brevini, Elena F M Manzoni, Sergio Ledda, Fulvio Gandolfi
Cell remarkable ability to self-organize and rearrange in functional organoids has been greatly boosted by the recent advances in 3-D culture technologies and materials. This approach can be presently applied to model human organ development and function "in a dish" and to predict drug response in a patient specific fashion.Here we describe a protocol that allows for the derivation of functional pancreatic mini-organoids from skin biopsies. Cells are suspended in a drop of medium and encapsulated with hydrophobic polytetrafluoroethylene (PTFE) powder particles, to form microbioreactors defined as "Liquid Marbles," that stimulate cell coalescence and 3-D aggregation...
July 13, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28702884/decellularization-of-large-tendon-specimens-combination-of-manually-performed-freeze-thaw-cycles-and-detergent-treatment
#5
Susanne Pauline Roth, Ina Erbe, Janina Burk
Reliable decellularization techniques applicable to tendon tissue play a critical role in the field of current tissue engineering. Particularly, an application as three-dimensional culture model for in vitro research and translational approaches to establish graft-based tendon repair as a routine clinical tool represent two main application fields for decellularized tendon scaffolds. Considering methodological issues of tendon decellularization, one of the major challenges lies in the preservation of the tendon-specific extracellular matrix (ECM) architecture to reflect natural tissue characteristic as best as possible...
July 13, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28674937/scaffold-based-and-scaffold-free-testicular-organoids-from-primary-human-testicular-cells
#6
Yoni Baert, Charlotte Rombaut, Ellen Goossens
Organoid systems take advantage of the self-organizing capabilities of cells to create diverse multi-cellular tissue surrogates that constitute a powerful novel class of biological models. Clearly, the formation of a testicular organoid (TO) in which human spermatogenesis can proceed from a single-cell suspension would exert a tremendous impact on research and development, clinical treatment of infertility, and screening of potential drugs and toxic agents. Recently, we showed that primary adult and pubertal human testicular cells auto-assembled in TOs either with or without the support of a natural testis scaffold...
July 4, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28656583/decellularization-of-intact-lung-tissue-through-vasculature-and-airways-using-negative-and-positive-pressure
#7
Steven Skolasinski, Angela Panoskaltsis-Mortari
Decellularization allows the production of extracellular matrix (ECM) scaffolds. Here we describe the use of combined positive pressure and negative pressure to drive decellularization reagents into the vasculature and airways, respectively, of structurally intact lungs in order to remove cells and cellular material leaving an intact ECM scaffold.
June 28, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28560513/immunohistochemical-detection-of-sphingosine-1-phosphate-and-sphingosine-kinase-1-in-human-tissue-samples-and-cell-lines
#8
Gary M Reynolds, Barbara Visentin, Roger Sabbadini
Sphingosine-1-phosphate (S1P) and the enzyme primarily responsible for its production, sphingosine kinase-1 (SphK-1), are dysregulated in multiple human diseases including cancer, multiple sclerosis (MS), diabetes, neurological diseases, fibrosis, and certain pathologies associated with impaired angiogenesis such as age-related macular degeneration (AMD). Antibody-based techniques to identify and localize S1P and SphK-1 within cells and tissue specimens represent a powerful tool, not only to understand biological role of these molecules but also to validate these unique in-class targets in multiple state diseases...
May 31, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28550502/decellularization-methods-for-scaffold-fabrication
#9
Sweta K Gupta, Narayan C Mishra
Decellularization is the process of removal of native cells from tissue, leaving behind a three-dimensional (3D) ultrastructure of extracellular matrix (ECM) proteins while preserving the bioactivity and mechanics of the tissue. It offers a unique top-down approach for fabricating ECM based natural scaffold for tissue engineering application. Herein, this chapter presents the fabrication of decellularized scaffold employing different methods: whole organ perfusion, immersion and agitation, pressure gradient, and supercritical fluid...
May 27, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28540560/alveolar-rhabdomyosarcoma-decellularization
#10
Michela Pozzobon, Mattia Saggioro, Stefania D'Agostino, Gianni Bisogno, Maurizio Muraca, Piergiorgio Gamba
In cancer research, it is an urgent need in the obtainment of a simple and reproducible model that mimics in all the complexity the pathological microenvironment. Specifically, the will to improve the overall survival of young patients affected by rhabdomyosarcoma compels researchers to develop new models resembling the multifaceted environment of the pathology to deeply study the disease under novel and different aspects. To this end, we developed a decellularization protocol for alveolar rhabdomyosarcoma (ARMS) able to maintain the three-dimensional structure...
May 25, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28540559/ceramide-and-s1p-signaling-in-embryonic-stem-cell-differentiation
#11
Guanghu Wang, Stefka D Spassieva, Erhard Bieberich
Bioactive sphingolipids are important regulators for stem cell survival and differentiation. Most recently, we have coined the term "morphogenetic lipids" for sphingolipids that regulate stem cells during embryonic and postnatal development. The sphingolipid ceramide and its derivative, sphingosine-1-phosphate (S1P), can act synergistically as well as antagonistically on embryonic stem (ES) cell differentiation. We show here simple as well as state-of-the-art methods to analyze sphingolipids in differentiating ES cells and discuss new protocols to use ceramide and S1P analogs for the guided differentiation of mouse ES cells toward neuronal and glial lineage...
May 25, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28540558/an-improved-isoform-selective-assay-for-sphingosine-kinase-1-activity
#12
Melissa R Pitman, Lorena T Davies, Stuart M Pitson
Sphingosine kinases (SK) are the sole enzymes responsible for the production of sphingosine 1-phosphate (S1P). S1P is a signaling molecule with a plethora of targets, acting as both a second messenger intracellularly and extracellularly via a family of cell surface G-protein-coupled S1P receptors. The two sphingosine kinases, SK1 and SK2, arise from different genes and have some distinct and overlapping cellular functions that are regulated in part by differential cellular localization, developmental expression, and catalytic properties...
May 25, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28510115/application-of-decellularized-tissue-scaffolds-in-ovarian-tissue-transplantation
#13
Enes Taylan, Kutluk Oktay
In tissue engineering, decellularized scaffolds have been proved to have remarkable capacity to promote regeneration in various organs such as kidney, heart, lung, and liver. Marrying the field of cryobiology and reproductive medicine resulted in considerable progress and breakthroughs, which led to the emergence of ovarian tissue cryopreservation and transplantation as a promising option for fertility preservation. Here we describe an innovative application of decellularized tissue scaffolds as a regenerative platform for reconstruction of ovarian grafts for auto-transplantation...
May 17, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28497335/methods-for-analyzing-sphingosine-1-phosphate-signaling-in-human-and-mouse-primary-mast-cells
#14
Alena P Chumanevich, Piper A Wedman, Carole A Oskeritzian
Mast cells produce a potently bioactive sphingolipid metabolite sphingosine-1-phosphate (S1P) constitutively and upon activation. The ligation of S1P to its type 2 receptor on mast cells triggers a novel downstream signaling pathway that we discovered links activation of transcription factor signal transducer and activator of transcription 3 to mast cell-derived chemokine release in both humans and mice. In this chapter, we describe the methods used to study S1P signaling in human and mouse primary mast cells...
May 12, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28497334/short-overview
#15
Norihiko Furuya
Mitochondrial autophagy (mitophagy) is a mitochondrial quality control mechanism that selectively removes damaged mitochondria via autophagic degradation. Autophagic adaptor/receptor proteins contribute to the selective degradation of damaged mitochondria by autophagy. A part of them containing both ubiquitin binding domains and Atg8 interacting motif (AIM)/LC3 interacting region (LIR) motifs, which bind to the autophagy-related protein 8 (Atg8) family (LC3 and GABARAP family), lead ubiquitylated (damaged) mitochondria to selective removal...
May 12, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28493213/3d-printing-composite-polycaprolactone-decellularized-bone-matrix-scaffolds-for-bone-tissue-engineering-applications
#16
Alexandra N Rindone, Ethan Nyberg, Warren L Grayson
Millions of patients worldwide require bone grafts for treatment of large, critically sized bone defects from conditions such as trauma, cancer, and congenital defects. Tissue engineered (TE) bone grafts have the potential to provide a more effective treatment than current bone grafts since they would restore fully functional bone tissue in large defects. Most bone TE approaches involve a combination of stem cells with porous, biodegradable scaffolds that provide mechanical support and degrade gradually as bone tissue is regenerated by stem cells...
May 11, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28493212/composite-bioscaffolds-incorporating-decellularized-ecm-as-a-cell-instructive-component-within-hydrogels-as-in-vitro-models-and-cell-delivery-systems
#17
Arthi Shridhar, Elizabeth Gillies, Brian G Amsden, Lauren E Flynn
Decellularized tissues represent promising biomaterials, which harness the innate capacity of the tissue-specific extracellular matrix (ECM) to direct cell functions including stem cell proliferation and lineage-specific differentiation. However, bioscaffolds derived exclusively from decellularized ECM offer limited versatility in terms of tuning biomechanical properties, as well as cell-cell and cell-ECM interactions that are important mediators of the cellular response. As an alternative approach, in the current chapter we describe methods for incorporating cryo-milled decellularized tissues as a cell-instructive component within a hydrogel carrier designed to crosslink under mild conditions...
May 11, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28488244/decellularized-iliotibial-band-recolonized-with-allogenic-homotopic-fibroblasts-or-bone-marrow-derived-mesenchymal-stromal-cells
#18
Clemens Gögele, Silke Schwarz, Benjamin Ondruschka, Niels Hammer, Gundula Schulze-Tanzil
Decellularized scaffolds present promising biomimetic approaches in various fields of tissue engineering. Different tissues have been selected for decellularization, among them extracellular matrix (ECM)-rich tissues such as tendons, ligaments and cartilage. The dense ECM of ligaments is particularly challenging to achieve a completely non-immunogenic ECM void of any cells. Here, the methods for decellularization adapted to ligamentous tissue of the iliotibial band (ITB) are presented along with cell isolation and several recolonization techniques using allogenic ITB-derived fibroblasts or mesenchymal stromal cells (MSCs)...
May 10, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28488243/monitoring-of-iron-depletion-induced-mitophagy-in-pathogenic-yeast
#19
Koichi Tanabe, Minoru Nagi
Mitophagy, which is the degradation of mitochondria via selective autophagic machinery, is thought to be involved in regulating the mass and function of mitochondria. Methods for detection of mitophagy have been reported for several fungal cells including some budding yeast, methylotrophic yeast, and filamentous fungi. Mitophagy in Saccharomyces cerevisiae is activated under nitrogen-poor conditions; however, the regulatory mechanism of mitophagy in most fungi has not been elucidated. Here we describe methods to monitor mitophagy in the pathogenic yeast Candida glabrata under iron-depleted conditions but not under nitrogen starvation...
May 10, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28466262/observation-of-parkin-mediated-mitophagy-in-pancreatic-%C3%AE-cells
#20
Atsushi Hoshino, Satoaki Matoba
Mitophagy is a cellular process of autophagy-based mitochondrial degradation that eliminates dysfunctional mitochondria and ensures cellular homeostasis. In pancreatic islet β-cells, mitochondria play a pivotal role in glucose-stimulated insulin secretion through ATP production from glucose oxidation. Recent studies have shown that impaired mitophagy and the subsequent mitochondrial compromise contribute to β-cell dysfunction and glucose intolerance. In this chapter, we describe a protocol to monitor Parkin-mediated mitophagy in pancreatic MIN6 β-cells using flow cytometry and a pH-sensitive fluorophore, mKeima...
May 3, 2017: Methods in Molecular Biology
journal
journal
31173
1
2
Fetch more papers »
Fetching more papers... Fetching...
Read by QxMD. Sign in or create an account to discover new knowledge that matter to you.
Remove bar
Read by QxMD icon Read
×

Search Tips

Use Boolean operators: AND/OR

diabetic AND foot
diabetes OR diabetic

Exclude a word using the 'minus' sign

Virchow -triad

Use Parentheses

water AND (cup OR glass)

Add an asterisk (*) at end of a word to include word stems

Neuro* will search for Neurology, Neuroscientist, Neurological, and so on

Use quotes to search for an exact phrase

"primary prevention of cancer"
(heart or cardiac or cardio*) AND arrest -"American Heart Association"