Federal University of Rio de Janeiro

8 ARTICLES PUBLISHED IN JoVE

Chemistry

CN-GELFrEE - Clear Native Gel-eluted Liquid Fraction Entrapment Electrophoresis

Rafael D. Melani^*1,2, Henrique S. Seckler^*1, Owen S. Skinner¹, Luis H. F. Do Vale^1,3, Adam D. Catherman¹, Pierre C. Havugimana¹, Marcelo Valle de Sousa³, Gilberto B. Domont², Neil L. Kelleher¹, Philip D. Compton¹

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.

Bioengineering

Preparation and Characterization of Nanoliposomes for the Entrapment of Bioactive Hydrophilic Globular Proteins

Anna C. N. T. F. Corrêa¹, Patricia R. Pereira¹, Vânia M. F. Paschoalin¹

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.

Bioengineering

Generating a Fractal Microstructure of Laminin-111 to Signal to Cells

Camila Hochman-Mendez¹, Tatiana Coelho-Sampaio², Ariel J. Kent³, Jamie L. Inman³, Mina J. Bissell³, Claire Robertson⁴

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.

Environment

A Field Primer for Monitoring Benthic Ecosystems Using Structure-From-Motion Photogrammetry

Ty N. F. Roach¹, Shreya Yadav¹, Carlo Caruso¹, Jenna Dilworth^1,2, Catherine M. Foley¹, Joshua R. Hancock¹, Joel Huckeba¹, Ariana S. Huffmyer¹, Kira Hughes¹, Valerie A. Kahkejian¹, Elizabeth M.P. Madin¹, Shayle B. Matsuda¹, Michael McWilliam¹, Spencer Miller^1,3, Erika P. Santoro⁴, Mariana Rocha de Souza¹, Damaris Torres-Pullizaa¹, Crawford Drury¹, Joshua S. Madin¹

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.

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 Neto¹

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.

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.

Biology

Establishing a Diaphyseal Femur Fracture Model in Mice

Bianca Braga Frade^*1, Leonardo Dias da Cunha Muller^*2, Danielle Cabral Bonfim³

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.

Biology

Evaluation of Right Ventricular Function in Experimental Models of Pulmonary Arterial Hypertension

Renata Trabach Santos¹, Maria Eduarda de Sá Freire Onofre¹, Victória Marques Barbosa¹, Nazareth Novaes Rocha¹, Patricia Rieken Macedo Rocco¹, Fernanda Ferreira Cruz¹, Pedro Leme Silva¹

¹Laboratory of Pulmonary Investigation, Institute of Biophysics Carlos Chagas Filho, Federal University of Rio de Janeiro

The presentation of detailed methods for the evaluation of right ventricle function will enhance the quality and reliability of pulmonary hypertension research, offering a robust framework for future studies and enhancing reproducibility across different laboratories.