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Free University Berlin

3 ARTICLES PUBLISHED IN JoVE

Medicine

Patient Derived Cell Culture and Isolation of CD133⁺ Putative Cancer Stem Cells from Melanoma

Yvonne Welte^1,2, Cathrin Davies^1,3, Reinhold Schäfer^1,4, Christian R.A. Regenbrecht^1,3,4

¹Institute of Pathology, Laboratory of Molecular Tumor Pathology, Charité - Universitätsmedizin Berlin, ²Institute for Chemistry and Biochemistry, Free University Berlin, ³Laboratory for Functional Genomics Charité (LFGC), Charité - Universitätsmedizin Berlin, ⁴Comprehensive Cancer Center Charité, Charité - Universitätsmedizin Berlin

This article describes the preparation of freshly obtained melanoma tissue into primary cell cultures, and how to remove contaminations of erythrocytes and fibroblasts from the tumor cells. Finally, we describe how CD133⁺ putative melanoma stem cells are sorted from the CD133^- bulk using Magnetic Activated Cell Sorting (MACS).

Behavior

Systematic Assessment of Well-Being in Mice for Procedures Using General Anesthesia

Katharina Hohlbaum^1,2, Bettina Bert^2,3, Silke Dietze², Rupert Palme⁴, Heidrun Fink², Christa Thöne-Reineke¹

¹Institute of Animal Welfare, Animal Behavior and Laboratory Animal Science, Department of Veterinary Medicine, Freie Universität Berlin, ²Institute of Pharmacology and Toxicology, Department of Veterinary Medicine, Freie Universität Berlin, ³German Federal Institute for Risk Assessment (BfR), ⁴Unit of Physiology, Pathophysiology and Experimental Endocrinology, Department of Biomedical Sciences, University of Veterinary Medicine

We developed a protocol to assess well-being in mice during procedures using general anesthesia. A series of behavioral parameters indicating levels of well-being as well as glucocorticoid metabolites were analyzed. The protocol can serve as a general aid to estimate the degree of severity in a scientific, animal-centered manner.

Neuroscience

Noninvasive EEG Recordings from Freely Moving Piglets

Nora V. de Camp^1,2, Silke Dietze¹, Markus Klaßen³, Jürgen Bergeler¹

¹Institute of Animal Welfare, Animal Behavior and Laboratory Animal Science, Veterinary Medicine, Free University Berlin, ²Department of Biology, Behavioral Physiology, Humboldt University Berlin, ³Lehr- und Versuchsanstalt für Viehhaltung Hofgut Neumühle

Here, we present a protocol to record telemetric electroencephalograms (EEGs) from freely moving piglets directly in the pigpen without the use of a sedative, making it possible to record typical EEG patterns during non-REM sleep, like spindle bursts.