University of Edinburgh

8 ARTICLES PUBLISHED IN JoVE

JoVE Journal

Use of Human Perivascular Stem Cells for Bone Regeneration

Aaron W. James^*1, Janette N. Zara^*2, Mirko Corselli², Michael Chiang¹, Wei Yuan², Virginia Nguyen¹, Asal Askarinam¹, Raghav Goyal¹, Ronald K. Siu³, Victoria Scott¹, Min Lee³, Kang Ting¹, Bruno Péault^2,4, Chia Soo²

¹Dental and Craniofacial Research Institute and Section of Orthodontics, School of Dentistry, UCLA, ²UCLA and Orthopaedic Hospital, Department of Orthopaedic Surgery and the Orthopaedic Hospital Research Center, UCLA, ³Department of Bioengineering, UCLA, ⁴Center for Cardiovascular Science, University of Edinburgh

Human perivascular stem cells (PSCs) are a novel stem cell class for skeletal tissue regeneration similar to mesenchymal stem cells (MSCs). PSCs can be isolated by FACS (fluorescence activated cell sorting) from adipose tissue procured during standard liposuction procedures, then combined with an osteoinductive scaffold to achieve bone formation in vivo.

Neuroscience

Preparation of Parasagittal Slices for the Investigation of Dorsal-ventral Organization of the Rodent Medial Entorhinal Cortex

Hugh Pastoll¹, Melanie White², Matthew Nolan²

¹Neuroinformatics DTC, University of Edinburgh , ²Centre for Integrative Physiology, University of Edinburgh

We describe procedures for preparation and electrophysiological recording from brain slices that maintain the dorsal-ventral axis of the medial entorhinal cortex (MEC). Because neural encoding of location follows a dorsal-ventral organization within the MEC, these procedures facilitate investigation of cellular mechanisms important for navigation and memory.

Biology

Analysis of Single-cell Gene Transcription by RNA Fluorescent In Situ Hybridization (FISH)

Elena Ronander^1,2, Dominique C. Bengtsson^1,2, Louise Joergensen^1,2, Anja T. R. Jensen^1,2, David E. Arnot^1,2,3

¹Centre for Medical Parasitology, Department of International Health, Immunology & Microbiology, Faculty of Health Sciences, University of Copenhagen, ²Department of Infectious Diseases, Copenhagen University Hospital (Rigshospitalet), ³Institute of Infection and Immunology Research, School of Biology, University of Edinburgh

Fluorescent in situ hybridization (FISH) to identify mRNA transcripts in individual cells allows analysis of polygenic activity such as the simultaneous transcription of more than one member of the var multigene family in Plasmodium falciparum infected erythrocytes ¹. The technique is adaptable and can be used on different types of genes, cells and organisms.

Biology

Genomic Transformation of the Picoeukaryote Ostreococcus tauri

Gerben van Ooijen¹, Kirsten Knox¹, Katalin Kis¹, François-Yves Bouget^2,3, Andrew J. Millar¹

¹SynthSys, University of Edinburgh , ²Centre National de la Recherche Scientifique, Université Pierre et Marie Curie, Paris 06, ³UMR 7621, Laboratoire d'Océanographie Microbienne, Observatoire Océanologique, Banyuls-sur-Mer, Université Pierre et Marie Curie, Paris 06

This article describes genetic transformation of the unicellular marine alga Ostreococcus tauri by electroporation. This eukaryotic organism is an effective model platform for higher plants, possesing greatly reduced genomic and cellular complexity and being readily amenable to both cell culture and chemical biology.

Biology

Isolation of Blood-vessel-derived Multipotent Precursors from Human Skeletal Muscle

William C.W. Chen¹, Arman Saparov^2,3, Mirko Corselli⁴, Mihaela Crisan⁵, Bo Zheng⁶, Bruno Péault^7,8, Johnny Huard⁹

¹Stem Cell Research Center, Department of Bioengineering and Orthopedic Surgery, University of Pittsburgh, ²Department of Orthopedic Surgery, University of Pittsburgh, ³Nazarbayev University Research and Innovation System, Nazarbayev University, ⁴Department of Orthopaedic Surgery, UCLA Orthopaedic Hospital and the Orthopaedic Hospital Research Center, University of California at Los Angeles, ⁵Department of Cell Biology, Erasmus MC Stem Cell Institute, ⁶OHSU Center for Regenerative Medicine, Oregon Health & Science University, ⁷Centre for Cardiovascular Science and MRC Centre for Regenerative Medicine, Queen's Medical Research Institute and University of Edinburgh, ⁸David Geffen School of Medicine and the Orthopaedic Hospital Research Center, University of California at Los Angeles, ⁹Stem Cell Research Center, Department of Orthopedic Surgery and McGowan Institute for Regenerative Medicine, University of Pittsburgh

Blood vessels within human skeletal muscle harbor several multi-lineage precursor populations that are ideal for regenerative applications. This isolation method allows simultaneous purification of three multipotent precursor cell populations respectively from three structural layers of blood vessels: myogenic endothelial cells from intima, pericytes from media, and adventitial cells from adventitia.

Developmental Biology

Isolation of Perivascular Multipotent Precursor Cell Populations from Human Cardiac Tissue

James E. Baily¹, William C.W. Chen^2,3, Nusrat Khan⁴, Iain R. Murray⁴, Zaniah N. González Galofre⁴, Johnny Huard^5,6, Bruno Péault^4,7

¹Centre for Cardiovascular Science, Queen's Medical Research Institute, University of Edinburgh, ²Department of Bioengineering and Orthopaedic Surgery, University of Pittsburgh, ³Research Laboratory of Electronics and Department of Biological Engineering, Massachusetts Institute of Technology, ⁴MRC Centre for Regenerative Medicine, University of Edinburgh, ⁵Stem Cell Research Center, Department of Orthopaedic Surgery, University of Pittsburgh, ⁶Department of Orthopaedic Surgery, University of Texas Health Science Center at Houston, ⁷Department of Orthopaedic Surgery, UCLA Orthopaedic Hospital, David Geffen School of Medicine, University of California at Los Angeles

Human cardiac tissue harbours multipotent perivascular precursor cell populations that may be suitable for myocardial regeneration. The technique described here allows for the simultaneous isolation and purification of two multipotent stromal cell populations associated with native blood vessels, i.e. CD146⁺CD34^- pericytes and CD34⁺CD146^- adventitial cells, from the human myocardium.

Biology

Rapid Analysis of Circadian Phenotypes in Arabidopsis Protoplasts Transfected with a Luminescent Clock Reporter

Louise L. Hansen¹, Gerben van Ooijen¹

¹Institute for Molecular Plant Sciences, University of Edinburgh

The circadian clock regulates about a third of the Arabidopsis transcriptome, but the percentage of genes that feed back into timekeeping remains unknown. Here we visualize a method to rapidly assess circadian phenotypes in any mutant line of Arabidopsis using luminescent imaging of a circadian reporter transiently expressed in protoplasts.

Biology

Human Adipose Tissue Micro-fragmentation for Cell Phenotyping and Secretome Characterization

Bianca Vezzani^1,2, Mario Gomez-Salazar¹, Joan Casamitjana¹, Carlo Tremolada³, Bruno Péault^1,4

¹MRC Center for Regenerative Medicine, University of Edinburgh, ²Dept. of Morphology, Surgery and Experimental Medicine, Section of General Pathology, University of Ferrara, ³Italian Image Institute, ⁴Orthopaedic Hospital Research Center and Broad Stem Cell Research Center, David Geffen School of Medicine, University of California

Here, we present human adipose tissue enzyme-free micro-fragmentation using a closed system device. This new method allows the obtainment of sub-millimeter clusters of adipose tissue suitable for in vivo transplantation, in vitro culture, and further cell isolation and characterization.