Wissenschaftliche Veröffentlichungen

An dieser Stelle werden wir die wissenschaftlichen Veröffentlichungen zu den Forschungsergebnissen im Rahmen von M2Aind nennen.

Jan-Hinrich Rabe1,2, Denis A. Sammour1,2, Sandra Schulz1,2, Bogdan Munteanu1,2, Martina
Ott3, Katharina Ochs3,5, Peter Hohenberger4, Alexander Marx4, Michael Platten3,4,
Christiane A. Opitz5,6, Daniel S. Ory7 & Carsten Hopf1,2

1Center for Applied Research in Applied Biomedical Mass Spectrometry (ABIMAS), Mannheim University of Applied Sciences, Mannheim, Germany
2Institute of Medical Technology, Heidelberg University and Mannheim University of Applied Sciences, Mannheim, Germany
3German Cancer Consortium (DKTK) CCU Neuroimmunology and Brain Tumor Immunology, German Cancer Research Center (DKFZ), Heidelberg, Germany
4University Medical Center Mannheim of Heidelberg University, Mannheim, Germany
5Brain Cancer Metabolism Group, German Cancer Research Center (DKFZ), Heidelberg, Germany
6Department of Neurology and National Center of Tumor Diseases, University Hospital Heidelberg, Heidelberg, Germany
7Diabetic Cardiovascular Disease Center and Department of Medicine, Washington University School of Medicine

In: Scientific Reports 8, Article number: 313 (2018)

Abstract: Multimodal imaging combines complementary platforms for spatially resolved tissue analysis that are poised for application in life science and personalized medicine. Unlike established clinical in vivo multimodality imaging, automated workflows for in-depth multimodal molecular ex vivo tissue analysis that combine the speed and ease of spectroscopic imaging with molecular details provided by mass spectrometry imaging (MSI) are lagging behind. Here, we present an integrated approach that utilizes non-destructive Fourier transform infrared (FTIR) microscopy and matrix assisted laser desorption/ionization (MALDI) MSI for analysing single-slide tissue specimen. We show that FTIR microscopy can automatically guide high-resolution MSI data acquisition and interpretation without requiring prior histopathological tissue annotation, thus circumventing potential human-annotation-bias while achieving >90% reductions of data load and acquisition time. We apply FTIR imaging as an upstream modality to improve accuracy of tissue-morphology detection and to retrieve diagnostic molecular signatures in an automated, unbiased and spatially aware manner. We show the general applicability of multimodal FTIR-guided MALDI-MSI by demonstrating precise tumor localization in mouse brain bearing glioma xenografts and in human primary gastrointestinal stromal tumors. Finally, the presented multimodal tissue analysis method allows for morphology-sensitive lipid signature retrieval from brains of mice suffering from lipidosis caused by Niemann-Pick type C disease.

Julia Klicksa, Elena von Molitora, Torsten Ertongur-Fauthb, Rüdiger Rudolfc and Mathias Hafnerc

aMannheim University of Applied Sciences, Institute of Molecular and Cell Biology,
Mannheim, Germany
bBRAIN AG, Zwingenberg, Germany
cInstitute of Medical Technology, Heidelberg University, Heidelberg, Germany

In: Journal of Cellular Biotechnology 3 (2017) 21–39

Abstract: Skin fulfils a plethora of eminent physiological functions ranging from physical barrier over immunity shield to the interface mediating social interaction. Prone to several acquired and inherited diseases, skin is therefore a major target of pharmaceutical and cosmetic research. The lack of similarity between human and animal skin and rising ethical concerns in the use of animal models have driven the search for novel realistic three-dimensional skin models. This review provides a survey of contemporary skin models and compares them in terms of applicability, reliability, cost and complexity.

Yasmina Martí a,b,1, Elina Nürnberga,c,1, Sandra Horschitzb, Mathias Hafnerc, Patrick Schlossb, Andreas Meyer-Lindenbergb and Thorsten Laua

aCentral Institute of Mental Health, Hector Institute for Translational Brain Research, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany
bCentral Institute of Mental Health, Medical Faculty Mannheim, Heidelberg University, Department Psychiatry and Psychotherapy, Mannheim, Germany
cInstitute of Molecular and Cell Biology, Mannheim University of Applied Sciences, Mannheim, Germany

In: Journal of Cellular Biotechnology 3 (2017) 61–80

Abstract: Major pathomechanisms underlying neurodegenerative diseases, such as Parkinson’s Disease, are still not well understood. Induced human pluripotent and rodent embryonic stem cells provide powerful disease models to address neurodegeneration-inducing pathomechanisms on a molecular and cellular level. Our aim is to establish a refined protocol to generate healthy and patient donor stem cell-derived dopaminergic neurons to investigate neurodegenerative events in vitro.METHODSHuman healthy donor- and patient-derived induced pluripotent stem cells were differentiated into stable dopaminergic progenitor cell lines and further differentiated into dopaminergic neurons. Induced pluripotent stem cells, neuronal progenitors and terminally differentiated neurons were characterized by confocal laser microscopy-based immunofluorescence analysis, live cell imaging demonstrating dopamine transporter-specific uptake of a fluorescent substrate and transcriptome analysis. Based on our immunofluorescence analysis, dopaminergic differentiation approaches predominantly yield dopaminergic neurons and GFAP-expressing glial cells. We detected a small partition of GABAergic neurons, yet neither serotonergic nor glutamatergic neurons. Dopaminergic neurons were successfully stained for pre- and postsynaptic and mitochondrial markers. Live cell imaging experiments verified dopamine transporter-dependent uptake of the fluorescent monoamine transporter substrate ASP+. Human stem cell-derived dopaminergic neurons are a suitable cellular system for fluorescence-based experimental approaches to address neurodegenerative events in vitro.

Valeh Rustamova,b, Rüdiger Rudolfa,b, Vugar Yagubluc, Hella-Monika Kuhna, Mario Vitacolonnaa,1 and Mathias Hafnera,b,1

aInstitute of Molecular and Cell Biology, Faculty of Biotechnology, Mannheim University
of Applied Sciences, Mannheim, Germany
bInstitute of Medical Technology, Heidelberg University, Heidelberg, Germany
cDepartment of General, Visceral, Vascular, and Thoracic Surgery, Frankfurt Hoechst Hospital,
Frankfurt, Germany

In: Journal of Cellular Biotechnology 3 (2017) 41–50

Abstract: In recent years, many different methods were introduced for generation of 3D cell culture. However, many currently available three-dimensional techniques are not suitable for certain cell lines and sometimes showed a lack of reproducibility. Therefore, specific protocols for cell lines are needed. In this work, we demonstrate different methods of generating 3D cell culture for SCC4 tongue cancer cell line and discuss their applicability. Using three different methods, tumor spheroids were generated from SCC4 cells and cultured for 20 days. To investigate the influence of initial seeding density on spheroid morphology and size during long term culture, the same set of different cell numbers was used for each method. Using phase-contrast microscopy, spheroids were monitored until day 20 and their sizes were determined. We observed that spheroids were formed within 24 hours regardless of the method and initial cell density. Further, in all groups the spheroid size was maximal at day 2, followed by a decline until day 20. Spheroids remained stable until day 20 independent of initial seeding concentration in all groups. We have generated long-term culture spheroids of SCC4 cells. The size of the spheroids can be influenced by varying the initial cell seeding density until day 20. This may be useful if different sizes of spheroids are required, e.g. in hypoxia research.