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The University of Manchester

7 ARTICLES PUBLISHED IN JoVE

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Bioengineering

Optimizing Attachment of Human Mesenchymal Stem Cells on Poly(ε-caprolactone) Electrospun Yarns
Lucy A. Bosworth 1, Sarah R. Rathbone 1, Sarah H. Cartmell 1
1School of Materials, The University of Manchester

This article describes a range of set-ups for seeding human mesenchymal stem cells onto materials, in this case electrospun yarns, that do not cover the base of standard culture well plates in order to maximize and quantify the number of cells that initially attach compared to the known seeding density.

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Neuroscience

Dorsal Root Ganglia Neurons and Differentiated Adipose-derived Stem Cells: An In Vitro Co-culture Model to Study Peripheral Nerve Regeneration
Alba C. de Luca 1, Alessandro Faroni 2, Adam J. Reid 2,3
1Centre for Neuroprosthesis, EPFL | STI | IMT/IBI | LSBI, 2Blond McIndoe Research Laboratories, Institute of Inflammation & Repair, The University of Manchester, 3University Hospital of South Manchester

Dorsal root ganglia (DRG) are structures containing the sensory neurons of the peripheral nervous system. When dissociated, they can be co-cultured with SC-like adipose-derived stem cells (ASC), providing a valuable model to study in vitro nerve regeneration and myelination, mimicking the in vivo environment at the injury site.

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Chemistry

Determining the Chemical Composition of Corrosion Inhibitor/Metal Interfaces with XPS: Minimizing Post Immersion Oxidation
Monika S. Walczak 1, Perla Morales-Gil 1,2, Turia Belashehr 1, Kiran Kousar 1, Paulina Arellanes Lozada 1,3, Robert Lindsay 1
1Corrosion and Protection Centre, School of Materials, The University of Manchester, 2Laboratorio de Caracterización de Materiales Sintéticos y Naturales, Instituto Mexicano del Petróleo, 3Departamento de Metalurgia y Materiales, Instituto Politécnico Nacional

A protocol to avoid the oxidation of metallic substrates during sample transfer from an inhibited acidic solution to an X-ray photoelectron spectrometer is presented.

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Neuroscience

Light-Induced Molecular Adsorption of Proteins Using the PRIMO System for Micro-Patterning to Study Cell Responses to Extracellular Matrix Proteins
Cristina Melero *1, Aljona Kolmogorova *1, Paul Atherton 1, Brian Derby 2, Adam Reid 3,4, Karin Jansen 5, Christoph Ballestrem 1
1Faculty of Biology, Medicine and Health. Division of Cell Matrix, Biology and Regenerative Medicine, Wellcome Trust Centre for Cell-Matrix Research, The University of Manchester, 2School of Materials, The University of Manchester, 3Blond McIndoe Laboratories, School of Biological Sciences, Faculty of Biology, Medicine and Health, Division of Cell Matrix Biology and Regenerative Medicine, The University of Manchester, Manchester Academic Health Science Centre, 4Department of Plastic Surgery & Burns, Wythenshawe Hospital, Manchester University NHS Foundation Trust, Manchester Academic Health Science Centre, 5Department of Pathology, UMC Utrecht

Our overall aim is to understand how cells sense extracellular cues that lead to directed axonal growth. Here, we describe the methodology of Light-Induced Molecular Adsorption of Proteins, used to produce defined micro-patterns of extracellular matrix components in order to study specific events that govern axon outgrowth and pathfinding.

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Genetics

Measuring Microbial Mutation Rates with the Fluctuation Assay
Rok Krašovec 1, Huw Richards 2,3, Guillaume Gomez 3, Danna R. Gifford 1, Adrien Mazoyer 4, Christopher G. Knight 3
1School of Biological Sciences, The University of Manchester, 2School of Science, University of Waikato, 3School of Natural Sciences, The University of Manchester, 4Department of Mathematics, Université du Québec à Montréal

Here, a protocol is presented to perform a fluctuation assay and estimate microbial mutation rate using phenotypic markers. This protocol will enable researchers to assay mutations in diverse microbes and environments, determining how genotype and ecological context affect spontaneous mutation rates.

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JoVE Core

Finite Element Modelling of a Cellular Electric Microenvironment
Miruna Verdes 1, Catherine Disney 1, Chinnawich Phamornnak 1, Lee Margetts 2, Sarah Cartmell 1,3
1Department of Materials, Faculty of Science and Engineering, The University of Manchester, 2Department of Mechanical, Aerospace and Civil Engineering, Faculty of Science and Engineering, The University of Manchester, 3The Henry Royce Institute, Royce Hub Building, The University of Manchester

This paper presents a strategy for building finite element models of fibrous conductive materials exposed to an electric field (EF). The models can be used to estimate the electrical input that cells seeded in such materials receive and assess the impact of changing the scaffold's constituent material properties, structure or orientation.

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Biology

Adaptation at the Extremes of Life: Experimental Evolution with the Extremophile Archaeon Sulfolobus acidocaldarius
Zahraa Al-Baqsami *1,2,3, Rebecca Lowry Palmer *1,3, Gwyneth Darwent 1, Andrew J. McBain 2, Christopher G. Knight 3, Danna R. Gifford 1
1Division of Evolution, Infection and Genomics, School of Biological Sciences, The University of Manchester, 2Division of Pharmacy and Optometry, School of Health Sciences, The University of Manchester, 3Department of Earth and Environmental Sciences, School of Natural Sciences, The University of Manchester

Here, we present an experimental evolution protocol for adaptation in thermophiles utilizing low-cost, energy-efficient bench-top thermomixers as incubators. The technique is demonstrated through the characterization of temperature adaptation in Sulfolobus acidocaldarius, an archaeon with an optimal growth temperature of 75 °C.

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