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School of Medicine University of Rostock

4 ARTICLES PUBLISHED IN JoVE

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Bioengineering

Cultivation of Human Neural Progenitor Cells in a 3-dimensional Self-assembling Peptide Hydrogel
Andrea Liedmann 1, Arndt Rolfs 1, Moritz J. Frech 1
1Albrecht-Kossel-Institute for Neuroregeneration, University of Rostock

Here we describe the use of a self-assembling 3-dimensional scaffold to culture human neural progenitor cells. We present a protocol to release the cells from the scaffolds to be analysed subsequently e.g. by flow cytometry. This protocol might be adapted to other cell types to perform detailed mechanistically studies.

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Biology

Gene Transfer into Older Chicken Embryos by ex ovo Electroporation
Jiankai Luo 1, Xin Yan 1, Juntang Lin 2, Arndt Rolfs 1
1Albrecht-Kossel-Institute for Neuroregeneration, School of Medicine University of Rostock, 2Institute of Anatomy I, School of Medicine University of Jena

A method of gene transfer into chicken embryos at later incubation stages (older than Hamburger and Hamilton stage (HH) 22) is described. This method overcomes disadvantages of in ovo electroporation applied to older chicken embryos and is a useful technique to study gene function and regulation at older developmental stages.

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Medicine

In Vitro Enzyme Measurement to Test Pharmacological Chaperone Responsiveness in Fabry and Pompe Disease
Jan Lukas 1, Anne-Marie Knospe 1, Susanne Seemann 1, Valentina Citro 2, Maria V. Cubellis 2, Arndt Rolfs 1,3
1Albrecht-Kossel-Institute, University Rostock Medical Center, 2Department of Biology, University Federico II, 3Centogene AG

There is a demand to make pre-clinical testing for a novel class of "orphan" drugs called pharmacological chaperones reproducible, fast, and efficient. We developed a simple, highly standardized, and versatile cell culture-based assay to screen for eligible patients as well as novel pharmacological chaperone drugs.

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Immunology and Infection

Generation of Monoclonal Antibodies Against Natural Products
Yue Zhang 1, Peng Cao 2, Fang Lu 3, Xin Yan 4, Bingqian Jiang 3, Jinjun Cheng 3, Huihua Qu 5
1School of Life Science, Beijing University of Chinese Medicine, 2Third Affiliated Hospital, Beijing University of Chinese Medicine, 3School of Basic Medical Sciences, Beijing University of Chinese Medicine, 4School of Chinese Materia Medica, Beijing University of Chinese Medicine, 5Center of Scientific Experiment, Beijing University of Chinese Medicine

This article provides a detailed protocol for the preparation and evaluation of monoclonal antibodies against natural products for use in various immunoassays. This procedure includes immunization, cell fusion, indirect competitive ELISA for positive clone screening, and monoclonal hybridoma preparation. The specifications for antibody characterization using MALDI-TOF-MS and ELISA analyses are also provided.

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