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Albert Einstein College of Medicine

33 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Passive Administration of Monoclonal Antibodies Against H. capsulatum and Others Fungal Pathogens
Allan J. Guimarães 1, Luis R. Martinez 1, Joshua D. Nosanchuk 1
1Department of Microbiology and Immunology, Albert Einstein College of Medicine

C57BL/6 mice have been used to study Hc pathogenesis and provide the best model. We are exploring the potential benefits of humoral immunity against this fungus and generated several mAbs [to histone H2B and a heat shock protein 60kDa] that we tested for their protective efficacy after intraperitoneal administration.

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Biology

Preparation of Adult Drosophila Eyes for Thin Sectioning and Microscopic Analysis
Andreas Jenny 1
1Department of Developmental and Molecular Biology, Albert Einstein College of Medicine

A standard approach to prepare adult Drosophila eyes for semi-thin sectioning and light microscopic analysis is presented here. The protocol can be used for gross morphological analysis of eye defects, or with the indicated adjustments can be used to determine genetic requirements of genes in specific cell types of the eye (e.g. clonal analysis of photoreceptors) or for electron microscopic analysis.

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Biology

Monitoring Equilibrium Changes in RNA Structure by 'Peroxidative' and 'Oxidative' Hydroxyl Radical Footprinting
Ravichandra Bachu *1, Frances-Camille S. Padlan *2, Sara Rouhanifard 2, Michael Brenowitz 2, Jörg C. Schlatterer 2
1Department of Chemistry, Hunter College , 2Department of Biochemistry, Albert Einstein College of Medicine

This protocol describes how to quantify the Mg(II)-dependent formation of RNA tertiary structure by two methods of hydroxyl radical footprinting.

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Biology

Real Time Analysis of Metabolic Profile in Ex Vivo Mouse Intestinal Crypt Organoid Cultures
Tuba Bas 1, Leonard H. Augenlicht 1,2
1Department of Medicine, Albert Einstein College of Medicine, 2Department of Cell Biology, Albert Einstein College of Medicine

Small intestinal crypt organoids cultured ex vivo provide a tissue culture system that recapitulates growth of crypts dependent on stem cells and their niche. We established a method to assay the metabolic profile in real time in primary mouse crypt organoids. We found organoids maintain physiological properties defined by their source.

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Immunology and Infection

Visualizing Non-lytic Exocytosis of Cryptococcus neoformans from Macrophages Using Digital Light Microscopy
Sabriya Stukes 1, Arturo Casadevall 1
1Department of Microbiology and Immunology, Albert Einstein College of Medicine

We describe how to visualize macrophage-C. neoformans (Cn) interactions in real time, with specific emphasis on the process of non-lytic exocytosis using digital light microscopy. Using this technique individually infected macrophages can be studied to ascertain various aspects of this phenomenon.

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Immunology and Infection

An Efficient and High Yield Method for Isolation of Mouse Dendritic Cell Subsets
Pooja Arora 1, Steven A. Porcelli 1,2
1Department of Microbiology and Immunology, Albert Einstein College of Medicine, 2Department of Medicine (Rheumatology), Albert Einstein College of Medicine

Distinct dendritic cell subsets exist as rare populations in lymphoid organs, and therefore are challenging to isolate in sufficient numbers and purity for immunological experiments. Here we describe a high efficiency, high yield method for isolation of all of the currently known major subsets of mouse splenic dendritic cells.

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Cancer Research

Coculture Assays to Study Macrophage and Microglia Stimulation of Glioblastoma Invasion
Salvatore Coniglio 1, Ian Miller 2, Marc Symons 3, Jeffrey E. Segall 4
1New Jersey Center for Science, Technology and Mathematics, Kean University, 2Department of Physiology and Medical Physics, Royal College of Surgeons in Ireland, 3The Feinstein Institute for Medical Research at North Shore-LIJ, 4Department of Anatomy and Structural Biology, Gruss Lipper Biophotonics Center, Albert Einstein College of Medicine

Understanding the malignant behavior of cancer requires creating accurate models of how tumor cells interact with components of the tumor microenvironment, such as macrophages. Here we describe two methods to study glioblastoma cell interaction with tumor associated macrophages and microglia where the effect on glioblastoma invasion is assessed.

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Medicine

Extended Time-lapse Intravital Imaging of Real-time Multicellular Dynamics in the Tumor Microenvironment
Allison S. Harney 1,2,3,4, Yarong Wang 1,3, John S. Condeelis 1,3,4, David Entenberg 1,3,4
1Department of Anatomy & Structural Biology, Albert Einstein College of Medicine, 2Department of Radiology, Albert Einstein College of Medicine, 3Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, 4Integrated Imaging Program, Albert Einstein College of Medicine

This protocol describes the use of multiphoton microscopy to perform extended time-lapse imaging of multicellular interactions in real time, in vivo at single cell resolution.

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Biology

Complete Workflow for Analysis of Histone Post-translational Modifications Using Bottom-up Mass Spectrometry: From Histone Extraction to Data Analysis
Simone Sidoli 1, Natarajan V. Bhanu 1, Kelly R. Karch 1, Xiaoshi Wang 1, Benjamin A. Garcia 1
1Epigenetics Program, Department of Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania

This protocol outlines a fully integrated workflow for characterizing histone post-translational modifications using mass spectrometry (MS). The workflow includes histone purification from cell cultures or tissues, histone derivatization and digestion, MS analysis using nano-flow liquid chromatography and instructions for data analysis. The protocol is designed for completion within 2 - 3 days.

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Cancer Research

Long-term High-Resolution Intravital Microscopy in the Lung with a Vacuum Stabilized Imaging Window
Carolina Rodriguez-Tirado 1, Takanori Kitamura 5, Yu Kato 1,2, Jeffery W. Pollard 1,2,5, John S. Condeelis 3,4, David Entenberg 3,4
1Department of Developmental and Molecular Biology, Albert Einstein College of Medicine, 2Department of Obstetrics/Gynecology and Woman’s Health, Albert Einstein College of Medicine, 3Department of Anatomy & Structural Biology, Albert Einstein College of Medicine, 4Gruss-Lipper Biophotonics Center Integrated Imaging Program, Albert Einstein College of Medicine, 5Medical Research Council Centre for Reproductive Health, Queen’s Medical Research Institute, University of Edinburgh

This protocol describes the use of multiphoton microscopy to perform long-term high-resolution, single cell imaging of the intact lung in real time using a vacuum stabilized imaging window.

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Biochemistry

Sample Preparation for Mass Spectrometry-based Identification of RNA-binding Regions
Robert Warneford-Thomson 1,4, Chongsheng He 1,2, Simone Sidoli 1,3, Benjamin A. Garcia 1,3, Roberto Bonasio 1,2
1Epigenetics Institute, University of Pennsylvania Perelman School of Medicine, 2Department of Cell and Developmental Biology, University of Pennsylvania Perelman School of Medicine, 3Department of Biochemistry and Biophysics, University of Pennsylvania Perelman School of Medicine, 4Graduate Group in Biochemistry and Biophysics, University of Pennsylvania Perelman School of Medicine

We describe a protocol to identify RNA-binding proteins and map their RNA-binding regions in live cells using UV-mediated photocrosslinking and mass spectrometry.

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Neuroscience

An In Vivo Duo-color Method for Imaging Vascular Dynamics Following Contusive Spinal Cord Injury
Chen Chen 1,2, Yi Ping Zhang 3, Yan Sun 1,4, Wenhui Xiong 1, Lisa B. E. Shields 3, Christopher B. Shields 3,5, Xiaoming Jin 1, Xiao-Ming Xu 1
1Spinal Cord and Brain Injury Research Group, Stark Neurosciences Research Institute, and Department of Neurological Surgery, Indiana University School of Medicine, 2Program in Medical Neuroscience, Stark Neurosciences Research Institute, Indiana University School of Medicine, 3Norton Neuroscience Institute, Norton Healthcare, 4Department of Human Anatomy & Histoembryology, School of Basic Medical Sciences, Fudan University, 5Department of Neurological Surgery, University of Louisville School of Medicine

We introduce an in vivo imaging method using two different fluorescent dyes to track dynamic spinal vascular changes following a contusive spinal cord injury in adult Sprague-Dawley rats.

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Bioengineering

Synthesis of Functionalized 10-nm Polymer-coated Gold Particles for Endothelium Targeting and Drug Delivery
Ming J. Cheng 1, Priya Prabakaran 1, Rajiv Kumar 2,3, Srinivas Sridhar 1,2,3, Eno E. Ebong 1,4,5
1Department of Chemical Engineering, Northeastern University, 2Nanomedicine Science and Technology Center, Northeastern University, 3Department of Physics, Northeastern University, 4Departments of Bioengineering, Northeastern University, 5Department of Neuroscience, Albert Einstein College of Medicine

We describe a method of synthesizing biocompatible 10-nm gold nanoparticles, functionalized by coating poly-ethylene glycol onto the surface. These particles can be used in vitro and in vivo for delivering therapeutics to nanoscale cellular and extracellular spaces that are difficult to access with conventional nanoparticle sizes.

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Immunology and Infection

Automated Measurement of Cryptococcal Species Polysaccharide Capsule and Cell Body
Quigly Dragotakes 1, Arturo Casadevall 1
1Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health

This technique describes an automated batch image processor designed to measure polysaccharide capsule and body radii. While initially designed for Cryptococcus neoformans capsule measurements the automated image processor can also be applied to other contrast based detection of circular objects.

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Biology

Broth Microdilution In Vitro Screening: An Easy and Fast Method to Detect New Antifungal Compounds
Calliandra Maria de-Souza-Silva 1, Fernanda Guilhelmelli 1, Daniel Zamith-Miranda 2,3, Marco Antônio de Oliveira 1, Joshua Daniel Nosanchuk 2,3, Ildinete Silva-Pereira *1, Patrícia Albuquerque *1,4
1Laboratory of Molecular Biology, Department of Cellular Biology, Institute of Biological Sciences, University of Brasília, 2Department of Microbiology and Immunology, Albert Einstein College of Medicine, 3Division of Infectious Diseases, Department of Medicine, Albert Einstein College of Medicine, 4Faculty of Ceilândia, University of Brasília

An easy and adaptable broth microdilution method for screening antifungal compounds and extracts.

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Genetics

Retrospective MicroRNA Sequencing: Complementary DNA Library Preparation Protocol Using Formalin-fixed Paraffin-embedded RNA Specimens
Olivier Loudig 1,2,3, Christina Liu 1,2, Thomas Rohan 3, Iddo Z. Ben-Dov 4
1Department of Research, Hackensack University Medical Center, 2Department of Medical Sciences, Seton Hall University, 3Department of Epidemiology and Population Health, Albert Einstein College of Medicine, 4Department of Nephrology and Hypertension, Hadassah - Hebrew University Medical Center

Formalin-fixed paraffin-embedded specimens represent a valuable source of molecular biomarkers of human diseases. Here we present a laboratory-based cDNA library preparation protocol, initially designed with fresh frozen RNA, and optimized for the analysis of archived microRNAs from tissues stored up to 35 years.

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Medicine

Updated Technique for Reliable, Easy, and Tolerated Transcranial Electrical Stimulation Including Transcranial Direct Current Stimulation
Helen Borges 1, Alexandra Dufau 1,2, Bhaskar Paneri 1, Adam J. Woods 3, Helena Knotkova 4,5, Marom Bikson 1
1Department of Biomedical Engineering, The City College of New York, CUNY, 2Department of Clinical and Health Psychology, Center for Cognitive Aging and Memory, 3McKnight Brain Institute, University of Florida, 4MJHS Institute for Innovation in Palliative Care, 5Department of Family and Social Medicine, Albert Einstein College of Medicine

When administering transcranial direct current stimulation (tDCS), reproducible electrode preparation and placement are vital for a tolerated and effective session. The purpose of this article is to demonstrate updated modern setup procedures for the administration of tDCS and related transcranial electrical stimulation techniques, such as transcranial alternating current stimulation (tACS).

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Behavior

Using the Race Model Inequality to Quantify Behavioral Multisensory Integration Effects
Jeannette R. Mahoney 1, Joe Verghese 1,2
1Department of Neurology, Division of Cognitive & Motor Aging, Albert Einstein College of Medicine, 2Department of Medicine, Division of Geriatrics, Albert Einstein College of Medicine

The current study aims to provide a step-by-step tutorial for calculating the magnitude of multisensory integration effects in an effort to facilitate the production of translational research studies across diverse clinical populations.

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Cancer Research

Assessing Tumor Microenvironment of Metastasis Doorway-Mediated Vascular Permeability Associated with Cancer Cell Dissemination using Intravital Imaging and Fixed Tissue Analysis
George S. Karagiannis 1,2,3, Jessica M. Pastoriza 1,2,4, Lucia Borriello 1,2, Rojin Jafari 1,2, Anouchka Coste 1,2,4, John S. Condeelis 1,2,3,4, Maja H. Oktay 1,2,3,5, David Entenberg 1,2,3
1Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, 2Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, 3Integrated Imaging Program, Albert Einstein College of Medicine, 4Department of Surgery, Montefiore Medical Center, 5Department of Pathology, Montefiore Medical Center

We describe two methods for assessing transient vascular permeability associated with tumor microenvironment of metastasis (TMEM) doorway function and cancer cell intravasation using intravenous injection of high-molecular weight (155 kDa) dextran in mice. The methods include intravital imaging in live animals and fixed tissue analysis using immunofluorescence.

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Biology

Improving the Accuracy of Flow Cytometric Assessment of Mitochondrial Membrane Potential in Hematopoietic Stem and Progenitor Cells Through the Inhibition of Efflux Pumps
Claudia Morganti 1,2,3, Massimo Bonora 1,2,3, Keisuke Ito 1,2,3,4
1Ruth L. and David S. Gottesman Institute for Stem Cell and Regenerative Medicine Research, Albert Einstein College of Medicine, 2Departments of Cell Biology and Stem Cell Institute, Albert Einstein College of Medicine, 3Department of Medicine, Albert Einstein College of Medicine, 4Albert Einstein Cancer Center and Diabetes Research Center, Albert Einstein College of Medicine

Xenobiotic efflux pumps are highly active in hematopoietic stem and progenitor cells (HSPCs) and cause extrusion of TMRM, a mitochondrial membrane potential fluorescent dye. Here, we present a protocol to accurately measure mitochondrial membrane potential in HSPCs by TMRM in the presence of Verapamil, an efflux pump inhibitor.

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Neuroscience

Isolation and Culture of Oculomotor, Trochlear, and Spinal Motor Neurons from Prenatal Islmn:GFP Transgenic Mice
Ryosuke Fujiki 1,2,3,4,9, Joun Y. Lee 1,2,10, Julie A. Jurgens 1,2,3,7, Mary C. Whitman 2,5,6, Elizabeth C. Engle 1,2,3,4,5,6,7,8
1Department of Neurology, Boston Children's Hospital, 2FM Kirby Neurobiology Center, Boston Children's Hospital, 3Department of Neurology, Harvard Medical School, 4Medical Genetics Training Program, Harvard Medical School, 5Department of Ophthalmology, Boston Children's Hospital, 6Department of Ophthalmology, Harvard Medical School, 7Broad Institute of M.I.T. and Harvard, 8Howard Hughes Medical Institute, 9Department of Neurology, Kokura Memorial Hospital, 10Department of Genetics, Albert Einstein College of Medicine

This work presents a protocol to yield homogeneous cell cultures of primary oculomotor, trochlear, and spinal motor neurons. These cultures can be used for comparative analyses of the morphological, cellular, molecular, and electrophysiological characteristics of ocular and spinal motor neurons.

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Genetics

Isolation of Adipose Tissue Nuclei for Single-Cell Genomic Applications
Gabrielle J. Benitez 1, Kosaku Shinoda 1,2,3
1Departments of Medicine, Albert Einstein College of Medicine, 2Departments of Molecular Pharmacology, Albert Einstein College of Medicine, 3Fleischer Institute of Diabetes and Metabolism

This publication describes a protocol for the isolation of nuclei from mature adipocytes, purification by fluorescence-activated sorting, and single-cell level transcriptomics.

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Medicine

Real-Time Monitoring of Neurocritical Patients with Diffuse Optical Spectroscopies
Rodrigo Menezes Forti 1,2, Marilise Katsurayama 2,3, Giovani Grisotti Martins 1, Lenise Valler 2,3, Andrés Quiroga 1,2, Luiz Simioni 1, Julien Menko 4, Antonio L. E. Falcão 3, Li Min Li 2,5, Rickson C. Mesquita 1,2
1Institute of Physics, University of Campinas, 2Brazilian Institute of Neuroscience and Neurotechnology, 3Clinical Hospital, University of Campinas, 4Department of Emergency Medicine, Albert Einstein College of Medicine, 5School of Medical Sciences, University of Campinas

Presented here is a protocol for non-invasively monitoring cerebral hemodynamics of neurocritical patients in real-time and at the bedside using diffuse optics. Specifically, the proposed protocol uses a hybrid diffuse optical systems to detect and display real-time information on cerebral oxygenation, cerebral blood flow and cerebral metabolism.

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Developmental Biology

A 3-D Tail Explant Culture to Study Vertebrate Segmentation in Zebrafish
M. Fethullah Simsek 1,3, Ertugrul M. Özbudak 1,2,3
1Division of Developmental Biology, Cincinnati Children’s Hospital Medical Center, 2Department of Pediatrics, University of Cincinnati College of Medicine, 3Department of Genetics, Albert Einstein College of Medicine

Here, we present the protocol for 3-D tissue culture of the zebrafish posterior body axis, enabling live study of vertebrate segmentation. This explant model provides control over axis elongation, alteration of morphogen sources, and subcellular resolution tissue-level live imaging.

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Developmental Biology

Establishing a High Throughput Epidermal Spheroid Culture System to Model Keratinocyte Stem Cell Plasticity
Yvon Woappi 1,2, Geraldine Ezeka 3, Justin Vercellino 4, Sean M. Bloos 5, Kim E. Creek 6, Lucia Pirisi 1
1Department of Pathology, Microbiology and Immunology, University of South Carolina School of Medicine, 2Department of Dermatology, Brigham and Women's Hospital, Harvard Medical School, 3Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine Baltimore, 4Department of Pathology, Albert Einstein College of Medicine, 5Department of Pediatrics, Vanderbilt University Medical Center, 6Department of Drug Discovery and Biomedical Sciences, College of Pharmacy, University of South Carolina

Here we describe a protocol for the systematic cultivation of epidermal spheroids in 3D suspension culture. This protocol has wide-ranging applications for use in a variety of epithelial tissue types and for the modeling of several human diseases and conditions.

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Cancer Research

Analysis of Liver Microenvironment During Early Progression of Non-Alcoholic Fatty Liver Disease-Associated Hepatocellular Carcinoma in Zebrafish
Cassia Michael 1,2, Francisco Juan Martínez-Navarro 1,2, Sofia de Oliveira 1,2,3,4
1Department of Developmental and Molecular Biology, Albert Einstein College of Medicine, 2Department of Medicine (Hepatology), Albert Einstein College of Medicine, 3Einstein-Mount Sinai Diabetes Research Center, Albert Einstein College of Medicine, 4Marion Bessin Liver Research Center, Albert Einstein College of Medicine

Here, we present how to generate a non-alcoholic fatty liver disease (NAFLD)-associated Hepatocellular Carcinoma (HCC) zebrafish model to study the impact of cholesterol surplus on liver microenvironment and immune cell landscape.

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Medicine

Surgical Techniques to Optimize Ovarian Reserve during Laparoscopic Cystectomy for Ovarian Endometrioma
Kathryn Saturnino 1,2,3, Osaro Obanor 1,2,3, Cynthia Arvizo 4, Julian A. Gingold 2,3,5
1Department of Obstetrics & Gynecology and Women's Health, Montefiore Medical Center, 2Albert Einstein College of Medicine, 3Department of OB/GYN, 1400 Pelham Pkwy S, Jacobi Medical Center, 4Division of Minimally Invasive Gynecologic Surgery, Department of OB/GYN, Jacobi Medical Center, 5Division of Reproductive Endocrinology and Infertility, Department of Obstetrics & Gynecology and Women's Health, Montefiore Medical Center

This protocol presents techniques to laparoscopically excise ovarian endometrioma, to perform adhesiolysis with sparing electrosurgical application, and to employ intraoperative chromopertubation to assess for genital tract patency. This systematic approach will facilitate optimal endometriosis management, guide concomitant adnexal surgeries, and enhance post-surgical fertility outcomes.

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Cancer Research

A Permanent Window for Investigating Cancer Metastasis to the Lung
Lucia Borriello *1,2, Brian Traub *1,2,3, Anouchka Coste 1,2,3, Maja H. Oktay 1,2,4,5, David Entenberg 1,2,4
1Department of Anatomy and Structural Biology, Einstein College of Medicine / Montefiore Medical Center, 2Gruss-Lipper Biophotonics Center, Einstein College of Medicine / Montefiore Medical Center, 3Department of Surgery, Einstein College of Medicine / Montefiore Medical Center, 4Integrated Imaging Program, Einstein College of Medicine / Montefiore Medical Cente, 5Department of Pathology, Einstein College of Medicine / Montefiore Medical Center

Here, we present a protocol for the surgical implantation of a permanently indwelling optical window for the murine thorax, which enables high-resolution, intravital imaging of the lung. The permanence of the window makes it well-suited to the study of dynamic cellular processes in the lung, especially those that are slowly evolving, such as metastatic progression of disseminated tumor cells.

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Biology

Global Level Quantification of Histone Post-Translational Modifications in a 3D Cell Culture Model of Hepatic Tissue
Jazmine-Saskya N. Joseph-Chowdhury 1, Stephanie Stransky 1, Sarah Graff 1, Ronald Cutler 1, Dejauwne Young 1, Julie S. Kim 1, Carlos Madrid-Aliste 1, Jennifer T. Aguilan 1, Edward Nieves 1, Yan Sun 1, Edwin J. Yoo 1, Simone Sidoli 1
1Department of Biochemistry, Albert Einstein College of Medicine

This protocol outlines how a three-dimensional cell culture system can be used to model, treat, and analyze chromatin modifications in a near-physiological state.

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Bioengineering

A Fluorescent Intravital Imaging Approach to Study Load-Induced Calcium Signaling Dynamics in Mouse Osteocytes
Karl J. Lewis 1, James F. Boorman-Padgett 3, Macy Castaneda 2, David C. Spray 4, Mia M. Thi 5,6, Mitchell B. Schaffler 3
1Meinig School of Biomedical Engineering, Cornell University, 2Sibley School of Mechanical and Aerospace Engineering, Cornell University, 3Department of Biomedical Engineering, The City College of New York, 4Department of Neuroscience, Albert Einstein College of Medicine, 5Department of Orthopaedic Surgery, Albert Einstein College of Medicine, 6Department of Molecular Pharmacology, Albert Einstein College of Medicine

The current article describes performing an intravital imaging approach to observe mechanically induced calcium signaling of embedded osteocytes in vivo in real-time in response to tissue-level mechanical loading of the mouse third metatarsal.

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Biochemistry

Semi-Automated Phenotypic Analysis of Functional 3D Spheroid Cell Cultures
Stephanie Stransky 1, Dejauwne Young 1, Karoline Mikkelsen 2,3, Annemette Præstegaard Thulesen 2, Helle Sedighi Frandsen 3, Simone Sidoli 1
1Department of Biochemistry, Albert Einstein College of Medicine, 2Department of Biochemistry and Molecular Biology, University of Southern Denmark, 3CelVivo ApS

We present a protocol for growing high-reproducible spheroids and their phenotypic characterization using image capture and proteomics.

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Immunology and Infection

Generation of Monocyte-Derived Dendritic Cells with Differing Sialylated Phenotypes
Vanessa C. C. Luz 1,2, Zélia Silva 1,2, Patrícia Sobral 1,2,3, Ankit Tanwar 4,5, Rachel L. Paterson 6, Paula A. Videira 1,2,7
1Associate Laboratory i4HB - Institute for Health and Bioeconomy, NOVA School of Science and Technology, Universidade NOVA de Lisboa, 2UCIBIO - Applied Molecular Biosciences Unit, Department of Life Sciences, NOVA School of Science and Technology, Universidade NOVA de Lisboa, 3LAQV and REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa, 4Department of Microbiology and Immunology, Albert Einstein College of Medicine, 5Department of Oncology, Albert Einstein College of Medicine, 6Stemmatters, Biotecnologia e Medicina Regenerativa SA, Parque de Ciência e Tecnologia Avepark, Zona Industrial da Gandra, 7CDG & Allies - Professionals and Patient Associations International Network (CDG & Allies - PPAIN), Department of Life Sciences, NOVA School of Science and Technology, Universidade NOVA de Lisboa

A unique, comprehensive protocol to generate de-sialylated human monocyte-derived dendritic cells (mo-DCs) from isolated peripheral blood mononuclear cells (PBMCs) using a sialidase treatment is presented. Further, methods to assess the phenotypic and functional characterization of mo-DCs and evaluate how sialidase treatment improves the maturation level of mo-DCs are described.

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Bioengineering

In Vitro Model Integrating Substrate Stiffness and Flow to Study Endothelial Cell Responses
Mohammad Hamrangsekachaee 1, Yu Chen 1, Emily R. Tressler 2, Sidi A. Bencherif 1,2,3,4, Eno E. Ebong 1,2,5
1Chemical Engineering Department, Northeastern University, 2Bioengineering Department, Northeastern University, 3Laboratoire de BioMécanique et BioIngénierie (BMBI), UMR CNRS, Sorbonne Universités, Université de Technologie of Compiègne (UTC), 4Harvard John A. Paulson School of Engineering and Applied Sciences, Harvard University, 5Neuroscience Department, Albert Einstein College of Medicine

We synthesized and characterized a tunable gelatin-based substrate for culturing vascular endothelial cells (ECs) under relevant vascular flow conditions. This biomimetic surface replicates both physiological and pathological conditions, enabling the study of mechanical forces on EC behavior and advancing our understanding of vascular health and disease mechanisms.

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