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  1. No Access

    Protocol

    Mouse Skeletal Muscle Decellularization

    Natural acellular matrices obtained from decellularization procedures are biocompatible and non-immunogenic materials considered promising tools for regenerative medicine purposes. Before in vivo implantation,...

    Martina Piccoli, Caterina Trevisan, Edoardo Maghin, Chiara Franzin, Michela Pozzobon (2017)

  2. No Access

    Protocol

    3D Stacked Construct: A Novel Substitute for Corneal Tissue Engineering

    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 ...

    Shrestha Priyadarsini, Sarah E. Nicholas, Dimitrios Karamichos (2017)

  3. No Access

    Protocol

    A Protocol to Prepare Decellularized Stem Cell Matrix for Rejuvenation of Cell Expansion and Cartilage Regeneration

    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...

    Jingting Li, Ming Pei (2017)

  4. No Access

    Protocol

    Decellularization of Bovine Small Intestinal Submucosa

    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 cell...

    Mahmut Parmaksiz, Ayşe Eser Elçin, Yaşar Murat Elçin (2017)

  5. No Access

    Protocol

    Analysis of S1P Receptor Expression by Uterine Immune Cells Using Standardized Multi-parametric Flow Cytometry

    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 s...

    Jianhong Zhang, Annie Bang, Stephen J. Lye (2017)

  6. No Access

    Protocol

    Antibody Uptake Assay in the Embryonic Zebrafish Forebrain to Study Notch Signaling Dynamics in Neural Progenitor Cells In Vivo

    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 ...

    Kai Tong, Mahendra Wagle, Su Guo (2017)

  7. No Access

    Protocol

    MicroRNA Expression Profiling by PCR Array in 2D and 3D Differentiated Neural Culture Systems and Target Validation

    MicroRNAs (miRNAs) have been proven to regulate gene expression at post-transcriptional level and are emerging as strong mediators in neural fate determination (Ambros, Nature 431(7006):350–355, 2004). Here, w...

    Lara Stevanato, Caroline Hicks, Lavaniya Thanabalasundaram, John D. Sinden (2017)

  8. No Access

    Protocol

    Monitoring of Paternal Mitochondrial Degradation in Caenorhabditis elegans

    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 p...

    Miyuki Sato, Ken Sato (2017)

  9. No Access

    Protocol

    Probing the Dynamics of Plasma Membrane Glutamate Transporters in Real Time by Total Internal Fluorescence Reflection Microscopy

    Plasma membrane solute carriers (SLCs) mediate the transport of a variety of substrates across the plasma membranes. There is a great interest in understanding how they work and how their density at the plasma...

    Eliana S. Di Cairano, Paola Marciani, Stefania Moretti, Carla Perego (2017)

  10. No Access

    Protocol

    Induction of PINK1/Parkin-Mediated Mitophagy

    PINK1/Parkin mitophagy is a key mechanism to contribute mitochondrial quality control, and the defects are thought to be a cause of those Parkinson’s disease onsets. Upon loss of mitochondrial membrane potenti...

    Shigeto Sato, Norihiko Furuya (2017)

  11. No Access

    Protocol

    A Simple Method of Generating 3D Brain Organoids Using Standard Laboratory Equipment

    3D brain organoids are a powerful tool with prospective application for the study of neural development and disease. Here we describe the growth factor-free method of generating cerebral organoids from feeder-...

    Magdalena Sutcliffe, Madeline A. Lancaster (2017)

  12. No Access

    Protocol

    A Cleanup Method for Mass Spectrometric Analysis of Sphingosine- and Ceramide-1-Phosphate in Blood and Solid Tissue Using a Phosphate Capture Molecule

    Cleanup technology and mass spectrometric determination of sphingosine-1-phosphate (S1P) using a phosphate capture molecule are shown. The protocol is rapid, requires neither thin-layer chromatography nor liqu...

    Jun-ichi Morishige, Ryouhei Yamashita, Tamotsu Tanaka, Kiyoshi Satouchi (2017)

  13. No Access

    Protocol

    The Use of Correlative Light-Electron Microscopy (CLEM) to Study PINK1/Parkin-Mediated Mitophagy

    In this chapter we describe the use of correlative light-electron microscopy (CLEM) to study, in cultured cells, the turnover of damaged mitochondria by PINK1/Parkin-dependent mitophagy. CLEM combines the adva...

    Chieko Kishi-Itakura, Folma Buss (2017)

  14. No Access

    Protocol

    Human Intestinal Enteroids: New Models to Study Gastrointestinal Virus Infections

    Human rotavirus (HRV) and human norovirus (HuNoV) infections are recognized as the most common causes of epidemic and sporadic cases of gastroenteritis worldwide. The study of these two human gastrointestinal ...

    Winnie Y. Zou, Sarah E. Blutt, Sue E. Crawford, Khalil Ettayebi, Xi-Lei Zeng (2017)

  15. No Access

    Protocol

    Immunocytochemical Monitoring of PINK1/Parkin-Mediated Mitophagy in Cultured Cells

    Both PINK1 and parkin are the responsible genes (PARK6 and PARK2, respectively) for familial early-onset Parkinson’s disease (PD). Several lines of evidences have suggested that mitochondrial dysfunction would...

    Motoki Fujimaki, Shinji Saiki, Yukiko Sasazawa, Kei-Ichi Ishikawa, Yoko Imamichi (2017)

  16. No Access

    Protocol

    Sphingosine-1-Phosphate (S1P) Signaling in Neural Progenitors

    Sphingosine-1-phosphate (S1P) and its receptors are important in nervous system development. Reliable in vitro human model systems are needed to further define specific roles for S1P signaling in neural develo...

    Phillip Callihan, Mohammed Alqinyah, Shelley B. Hooks (2017)

  17. No Access

    Protocol

    Measuring Sphingosine-1-Phosphate: Protein Interactions with the Kinetic Exclusion Assay

    By directly detecting the ligand-free binding sites in a sample, the kinetic exclusion assay (KinExA®) provides a compelling alternative to SPR-based techniques for determining equilibrium dissociation constants ...

    Jonathan K. Fleming, Jonathan M. Wojciak (2017)

  18. No Access

    Protocol

    Investigation of Yeast Mitophagy with Fluorescence Microscopy and Western Blotting

    Selective clearance of superfluous or dysfunctional mitochondria is a fundamental process that depends on the autophagic membrane trafficking pathways found in many cell types. This catabolic event, called mit...

    Sachiyo Nagumo, Koji Okamoto (2017)

  19. No Access

    Protocol

    Exploring MicroRNAs on NIX-Dependent Mitophagy

    The dysregulation of autophagy is implicated in many pathological disorders including infections, aging, neurodegenerative diseases, and cancer. Autophagy can be precisely controlled both transcriptionally and...

    Wen Li, Hao Chen, Shupeng Li, Guanghong Lin, Du Feng (2017)

  20. No Access

    Protocol

    Intestinal Crypt Organoid: Isolation of Intestinal Stem Cells, In Vitro Culture, and Optical Observation

    The isolation and culture of intestinal stem cells (ISCs) was first demonstrated in the very recent decade with the identification of ISC marker Lgr5. The growth of ISCs into crypt organoids provides an in vit...

    Yun Chen, Chuan Li, Ya-Hui Tsai, Sheng-Hong Tseng (2017)

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