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Friedrich-Alexander-Universität Erlangen-Nürnberg

5 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Lineage-reprogramming of Pericyte-derived Cells of the Adult Human Brain into Induced Neurons
Marisa Karow 1, Christian Schichor 2, Ruth Beckervordersandforth 3, Benedikt Berninger 1,4
1Department of Physiological Genomics, Institute of Physiology, Ludwig Maximilians University Munich, 2Tumor Biology Lab, Neurosurgical Clinic, Ludwig-Maximilians University Munich, 3Institut für Biochemie, Emil-Fischer-Zentrum, Friedrich-Alexander-Universität Erlangen-Nürnberg, 4Institute of Physiological Chemistry and Focus Program Translational Neuroscience, Johannes Gutenberg University Mainz

Targeting brain-resident cells for direct lineage-reprogramming offers new perspectives for brain repair. Here we describe a protocol of how to prepare cultures enriched for brain-resident pericytes from the adult human cerebral cortex and convert these into induced neurons by retrovirus-mediated expression of the transcription factors Sox2 and Ascl1.

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Chemistry

Isolating Free Carbenes, their Mixed Dimers and Organic Radicals
Annette Grünwald 1, Stephen J. Goodner 1, Dominik Munz 1
1Department of Chemistry and Pharmacy, Friedrich-Alexander-Universität Erlangen-Nürnberg

We present protocols for the isolation of stable heterocyclic carbenes. The synthesis of a cyclic (alkyl)(amino) carbene (CAAC) and an N-heterocyclic carbene (NHC) is demonstrated using filter cannulas and Schlenk technique. We furthermore present the synthesis of the related oxygen-sensitive, electron-rich mixed “Wanzlick dimer” and the reduced stable organic radical.

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JoVE Core

Cryogenic Liquid Jets for High Repetition Rate Discovery Science
Chandra B. Curry *1,2, Christopher Schoenwaelder *1,3, Sebastian Goede 4, Jongjin B. Kim 1, Martin Rehwald 5,6, Franziska Treffert 1,7, Karl Zeil 5, Siegfried H. Glenzer 1, Maxence Gauthier 1
1SLAC National Accelerator Laboratory, 2University of Alberta, 3Friedrich-Alexander-Universität Erlangen-Nürnberg, 4European XFEL, 5Helmholtz-Zentrum Dresden-Rossendorf, 6Technische Universität Dresden, 7Technische Universität Darmstadt

This protocol presents the operation and principles of micron-scale cylindrical and planar cryogenic liquid jets. Until now, this system has been used as a high repetition rate target in laser-plasma experiments. Anticipated cross-disciplinary applications range from laboratory astrophysics to material science, and eventually next-generation particle accelerators.

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Biology

Isolation of Human Ventricular Cardiomyocytes from Vibratome-Cut Myocardial Slices
Dominik J. Fiegle 1, Tilmann Volk 1,2, Thomas Seidel 1,2
1Institute of Cellular and Molecular Physiology, Friedrich-Alexander University Erlangen-Nürnberg, 2Muscle Research Center Erlangen (MURCE), Friedrich-Alexander-Universität Erlangen-Nürnberg

Presented is a protocol for the isolation of human and animal ventricular cardiomyocytes from vibratome-cut myocardial slices. High yields of calcium-tolerant cells (up to 200 cells/mg) can be obtained from small amounts of tissue (<50 mg). The protocol is applicable to myocardium exposed to cold ischemia for up to 36 h.

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Medicine

Preparation of Human Myocardial Tissue for Long-Term Cultivation
Jules Hamers 1,2, Payel Sen 1,2, Daphne Merkus 1,2,3, Thomas Seidel 4,5, Kun Lu 1,6, Andreas Dendorfer 1,2
1Walter-Brendel-Centre of Experimental Medicine, University Hospital, LMU Munich, 2German Center for Cardiovascular Research (DZHK), Partner Site Munich, Munich Heart Alliance (MHA), 3Division of Experimental Cardiology, Department of Cardiology, Erasmus MC, University Medical Center Rotterdam, 4Institute of Cellular and Molecular Physiology, Friedrich-Alexander-Universität Erlangen-Nürnberg, 5Muscle Research Center Erlangen (MURCE), Friedrich-Alexander-Universität Erlangen-Nürnberg, 6Department of Cardiac Surgery, University Hospital, LMU Munich

We present a protocol for ex vivo cultivation of human ventricular myocardial tissue. It allows for detailed analysis of contraction force and kinetics, as well as the application of pre- and afterload to mimic the in vivo physiological environment more closely.

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