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Federal University of Rio de Janeiro

7 ARTICLES PUBLISHED IN JoVE

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Chemistry

CN-GELFrEE - Clear Native Gel-eluted Liquid Fraction Entrapment Electrophoresis
Rafael D. Melani *1,2, Henrique S. Seckler *1, Owen S. Skinner 1, Luis H. F. Do Vale 1,3, Adam D. Catherman 1, Pierre C. Havugimana 1, Marcelo Valle de Sousa 3, Gilberto B. Domont 2, Neil L. Kelleher 1, Philip D. Compton 1
1Departments of Chemistry and Molecular Biosciences, Chemistry of Life Processes Institute, Proteomics Center of Excellence, Robert H. Lurie Comprehensive Cancer Center, Northwestern University, 2Institute of Chemistry, Proteomics Unit, Federal University of Rio de Janeiro, 3Department of Cell Biology, Brazilian Center for Protein Research, Laboratory of Biochemistry and Protein Chemistry, University of Brasilia

This protocol describes how to prepare and perform clear native gel-eluted liquid fraction entrapment electrophoresis (CN-GELFrEE), a native separation technique for non-covalent biomolecular assemblies and proteins from heterogeneous samples that is compatible with various downstream protein analysis techniques.

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Bioengineering

Preparation and Characterization of Nanoliposomes for the Entrapment of Bioactive Hydrophilic Globular Proteins
Anna C. N. T. F. Corrêa 1, Patricia R. Pereira 1, Vânia M. F. Paschoalin 1
1Chemistry Institute, Federal University of Rio de Janeiro

This study describes classical hydration using the thin lipid film method for nanoliposome preparation followed by nanoparticle characterization. A 47 kDa-hydrophilic and globular protein, tarin, is successfully encapsulated as a strategy to improve stability, avoid fast clearance, and promote controlled release. The method can be adapted to hydrophobic molecules encapsulation.

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Bioengineering

Generating a Fractal Microstructure of Laminin-111 to Signal to Cells
Camila Hochman-Mendez 1, Tatiana Coelho-Sampaio 2, Ariel J. Kent 3, Jamie L. Inman 3, Mina J. Bissell 3, Claire Robertson 4
1Regenerative Medicine Research, Texas Heart Institute, 2Institute of Biomedical Sciences, Federal University of Rio de Janeiro, 3Biological Systems and Engineering, Lawrence Berkeley National Lab, 4Materials Engineering Division, Lawrence Livermore National Lab

We describe three methods to generate Ln1 polymers with fractal properties that signal to cells differently compared to unpolymerized Ln1.

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Environment

A Field Primer for Monitoring Benthic Ecosystems Using Structure-From-Motion Photogrammetry
Ty N. F. Roach 1, Shreya Yadav 1, Carlo Caruso 1, Jenna Dilworth 1,2, Catherine M. Foley 1, Joshua R. Hancock 1, Joel Huckeba 1, Ariana S. Huffmyer 1, Kira Hughes 1, Valerie A. Kahkejian 1, Elizabeth M.P. Madin 1, Shayle B. Matsuda 1, Michael McWilliam 1, Spencer Miller 1,3, Erika P. Santoro 4, Mariana Rocha de Souza 1, Damaris Torres-Pullizaa 1, Crawford Drury 1, Joshua S. Madin 1
1Hawai'i Institute of Marine Biology, University of Hawai'i Manoa, 2Rosenstiel School of Marine and Atmospheric Science, University of Miami, 3Hawai'i Pacific University, 4Federal University of Rio de Janeiro

We provide a detailed protocol for conducting underwater structure-from-motion photogrammetry surveys to generate 3D models and orthomosaics.

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Developmental Biology

A Label-free Xenograft Model for Investigating the Behavior of Human Stem Cell Spheroids in Chick Embryos
Ingrid Rosenburg Cordeiro 1,2, José Marques de Brito Neto 1
1Morphological Sciences Program, Biomedical Sciences Institute, Federal University of Rio de Janeiro, 2Department of Life Science and Technology, Tokyo Institute of Technology

Here, we describe a method to transplant and identify human cell spheroids into chick embryos. This xenograft model uses the embryonic microenvironment as a source of instructive signals to assay cell migration, differentiation, and tropism and is especially suited for the study of primary and/or heterogeneous cell populations.

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Bioengineering

Large-Scale, Automated Production of Adipose-Derived Stem Cell Spheroids for 3D Bioprinting
Gabriela S. Kronemberger 1,2,3, Guilherme A. S. C. Miranda 1,2,4, Taisnara I. G. Silva 1,2,4, Rosângela M. Gonçalves 1,2, José M. Granjeiro 2,3,4,5, Leandra S. Baptista 1,2,3,4
1Nucleus of Multidisciplinary Research in Biology (Numpex-Bio), Federal University of Rio de Janeiro, 2Laboratory of Tissue Bioengineering, National Institute of Metrology, Quality and Technology (Inmetro), 3Post-graduation Program of Translational Biomedicine (Biotrans), Unigranrio, 4Post-graduation Program in Biotechnology, National Institute of Metrology, Quality and Technology (Inmetro), 5Dental School, Fluminense Federal Fluminense

Here, we describe the large-scale production of adipose-derived stromal/stem cell (ASC) spheroids using an automated pipetting system to seed the cell suspension, thus ensuring homogeneity of spheroid size and shape. These ASC spheroids can be used as building blocks for 3D bioprinting approaches.

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Biology

Establishing a Diaphyseal Femur Fracture Model in Mice
Bianca Braga Frade *1, Leonardo Dias da Cunha Muller *2, Danielle Cabral Bonfim 3
1Postgraduate Program in Biological Sciences – Biophysics, Institute of Biophysics Carlos Chagas Filho, Federal University of Rio de Janeiro, 2Postgraduate Program in Surgical Sciences, Faculty of Medicine, Federal University of Rio de Janeiro, 3Institute of Biomedical Sciences, Federal University of Rio de Janeiro

This protocol describes a surgical procedure for the establishment of a diaphyseal fracture in the femur of mice, which is stabilized with an intramedullary wire, for fracture healing studies.

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