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Methods in Molecular Biology

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https://www.readbyqxmd.com/read/28510115/application-of-decellularized-tissue-scaffolds-in-ovarian-tissue-transplantation
#1
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
#2
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
#3
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
#4
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
#5
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
#6
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
#7
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
#8
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
https://www.readbyqxmd.com/read/28456953/extracellular-matrix-from-whole-porcine-heart-decellularization-for-cardiac-tissue-engineering
#9
Matthew J Hodgson, Christopher C Knutson, Nima Momtahan, Alonzo D Cook
Decellularization of whole porcine hearts followed by recellularization with fully differentiated cells made from patient-specific human induced pluripotent stem cells (iPSCs) may provide the ultimate solution for patients with heart failure. Decellularization is the process of completely disrupting all cells and removing the cellular components (e.g., antigenic proteins, lipids, DNA) from organic tissue, leaving only the extracellular matrix (ECM). The decellularization of porcine hearts can be accomplished in 24 h and results in 98% DNA removal with only 6 h of detergent exposure...
April 30, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28456952/preparation-of-scaffolds-from-decellularized-testicular-matrix
#10
Yoni Baert, Ellen Goossens
Biological scaffolds composed of extracellular matrix (ECM) are typically derived by processes that involve decellularization of tissues or organs. Here we describe a simple and robust methodology for the preparation of decellularized testicular matrix (DTM) scaffolds with minimal damage to the native three-dimensional structure and tissue-specific ECM components. Such DTM scaffolds can help to gain a better insight into the molecular composition and function of testicular ECM and to develop new tissue engineering approaches to treat various types of male fertility disorders...
April 30, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28456951/s1p-synergizes-with-wall-shear-stress-and-other-angiogenic-factors-to-induce-endothelial-cell-sprouting-responses
#11
Camille L Duran, Roland Kaunas, Kayla J Bayless
Angiogenesis is the process of new blood vessel growth from pre-existing structures. During sprout initiation, endothelial cells (ECs) are activated by pro-angiogenic factors to degrade the basement membrane, migrate into the surrounding matrix, and form structures that anastomose to connect neighboring vessels. Sphingosine 1-phosphate (S1P) is a biologically active lysosphingolipid that is secreted by platelets and promotes angiogenesis under normal and pathological conditions by acting on ECs. In addition to biochemical factors, the endothelium is continuously subjected to mechanical forces in the form of wall shear stress (WSS) from fluid forces...
April 30, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28456950/a-rapid-fluorescence-assay-for-measuring-sphingosine-1-phosphate-transporter-activity-in-erythrocytes
#12
Naoki Kobayashi, Tsuyoshi Nishi
Sphingosine-1-phosphate (S1P) is an intercellular signaling molecule that is present in the plasma and plays an important role in recruiting lymphocytes from the thymus and secondary lymphoid organs. Erythrocytes are the most abundant cells in the blood and substantially contribute to the S1P supply in the plasma by releasing intracellularly synthesized S1P via an S1P transporter. Thus, the S1P transporter in erythrocytes is a potential target for immuno-suppressing drugs.In this chapter, we describe a rapid method for measuring the activity of the erythrocyte S1P transporter by using the fluorescent S1P analog, 7-nitro-2-1,3-benzoxadiazol-4-yl (NBD)-labeled S1P...
April 30, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28456949/monitoring-of-atg5-independent-mitophagy
#13
Satoko Arakawa, Shinya Honda, Satoru Torii, Masatsune Tsujioka, Shigeomi Shimizu
Mitophagy is a mitochondrial quality control mechanism where damaged and surplus mitochondria are removed by autophagy. There are at least two different mammalian autophagy pathways: the Atg5-dependent conventional pathway and an Atg5-independent alternative pathway; the latter is involved in the erythrocyte mitophagy. In this chapter we describe the various experimental approaches to assess Atg5-indepedndent mitophagy in mammalian cells.
April 30, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28451997/decellularization-of-bovine-small-intestinal-submucosa
#14
Mahmut Parmaksiz, Ayşe Eser Elçin, Yaşar Murat Elçin
Decellularization technology promises to overcome some of the significant limitations in the regenerative medicine field by providing functional biocompatible grafts. The technique involves removal of the cells from the biological tissues or organs for further use in tissue engineering and clinical interventions. There are significant differences between decellularization protocols due to the intrinsic properties of different tissue types and purpose of use. This multistep, chemical-solution-based protocol is optimized for the preparation of decellularized bovine small intestinal submucosa (SIS)...
April 28, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28451996/mouse-skeletal-muscle-decellularization
#15
Martina Piccoli, Caterina Trevisan, Edoardo Maghin, Chiara Franzin, Michela Pozzobon
Natural acellular matrices obtained from decellularization procedures are biocompatible and non-immunogenic materials considered promising tools for regenerative medicine purposes. Before in vivo implantation, these matrices must be efficiently decellularized, removing all the cellular components to avoid any immunogenic reaction. At the same time, it is important to maintain the original three-dimensional structure of the specific tissue. Here we describe a method: (1) to decellularize mouse quadriceps using a detergent-enzymatic treatment (DET) and (2) to assess decellularization efficiency and scaffold properties...
April 28, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28451995/a-protocol-to-prepare-decellularized-stem-cell-matrix-for-rejuvenation-of-cell-expansion-and-cartilage-regeneration
#16
Jingting Li, Ming Pei
Traditional ex vivo expansion of adult stem cells yields an insufficient quantity of less potent cells. Here we describe the fabrication of decellularized matrix deposited by synovium-derived stem cells (SDSCs). This matrix could serve as a three-dimensional expansion system to rejuvenate cells for proliferation and tissue-specific differentiation potential, which could benefit cartilage regeneration. The decellularized stem cell matrix (DSCM) might be a powerful system for tissue engineering and regeneration...
April 28, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28451994/3d-stacked-construct-a-novel-substitute-for-corneal-tissue-engineering
#17
Shrestha Priyadarsini, Sarah E Nicholas, Dimitrios Karamichos
Corneal trauma/injury often results in serious complications including permanent vision loss or loss of visual acuity which demands corneal transplantations or treatment with allogenic graft tissues. There is currently a huge shortage of donor tissue worldwide and the need for human corneal equivalents increases annually. In order to meet such demand the current clinical approach of treating corneal injuries is limited and involves synthetic and allogenic materials which have various shortcomings when it comes to actual transplantations...
April 28, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28434167/analysis-of-s1p-receptor-expression-by-uterine-immune-cells-using-standardized-multi-parametric-flow-cytometry
#18
Jianhong Zhang, Annie Bang, Stephen J Lye
Flow cytometry is a powerful tool for phenotypic and functional analyses of single immune cells. The increasing capability of flow cytometry technology has driven a more detailed understanding of immune cell subsets and functions in complex cellular systems such as the developing human decidua/placenta. We propose a standardized procedure for the isolation and analysis of human decidual natural killer (dNK) cells and this method can be extended to investigation of other uterine lymphocytes. Here this platform is used to examine the expression of sphingosine-1-phosphate (S1P) receptor and functional growth factors by dNK cells...
April 23, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28434166/antibody-uptake-assay-in-the-embryonic-zebrafish-forebrain-to-study-notch-signaling-dynamics-in-neural-progenitor-cells-in-vivo
#19
Kai Tong, Mahendra Wagle, Su Guo
Stem cells can generate cell fate heterogeneity through asymmetric cell division (ACD). ACD derives from the asymmetric segregation of fate-determining molecules and/or organelles in the dividing cell. Radial glia in the embryonic zebrafish forebrain are an excellent model for studying the molecular mechanisms regulating ACD of stem cells in vertebrates, especially for live imaging concerning in vivo molecular and cellular dynamics. Due to the current difficulty in expressing fluorescent reporter-tagged proteins at physiological levels in zebrafish for live imaging, we have developed an antibody uptake assay to label proteins in live embryonic zebrafish forebrain with high specificity...
April 23, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28425080/monitoring-of-paternal-mitochondrial-degradation-in-caenorhabditis-elegans
#20
Miyuki Sato, Ken Sato
In Caenorhabditis elegans embryos, paternally inherited mitochondria and their mitochondrial DNA are degraded via selective autophagy called allophagy (allogeneic organelle autophagy). This is a developmentally programmed autophagy and combined with C. elegans genetics and in vivo imaging provides a unique opportunity to analyze selective autophagy under physiological conditions.
April 20, 2017: Methods in Molecular Biology
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