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Establishing a human neuronal stem cell derived culture model to study neurodegeneration and neuroregeneration in multiple sclerosis
Author(s): ,
M. Alisch
Affiliations:
Max-Delbrück-Center for Molecular Medicine and Charité–Universitätsmedizin Berlin, Berlin, Germany
,
J. Kerkering
Affiliations:
Max-Delbrück-Center for Molecular Medicine and Charité–Universitätsmedizin Berlin, Berlin, Germany
,
L. Meyer-Arndt
Affiliations:
Max-Delbrück-Center for Molecular Medicine and Charité–Universitätsmedizin Berlin, Berlin, Germany
,
T. Crowley
Affiliations:
Max-Delbrück-Center for Molecular Medicine and Charité–Universitätsmedizin Berlin, Berlin, Germany
,
K. Rosiewicz
Affiliations:
Max-Delbrück-Center for Molecular Medicine and Charité–Universitätsmedizin Berlin, Berlin, Germany
V. Siffrin
Affiliations:
Max-Delbrück-Center for Molecular Medicine and Charité–Universitätsmedizin Berlin, Berlin, Germany
ECTRIMS Online Library. Alisch M. Oct 12, 2018; 228904
Marlen Alisch
Marlen Alisch
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Abstract: P1063

Type: Poster Sessions

Abstract Category: Pathology and pathogenesis of MS - Experimental models

The disease course of the chronic inflammatory and neurodegenerative autoimmune disease multiple sclerosis (MS) varies between benign forms and those with continuously increasing disability. The causes of this heterogeneity of disease course in between patients is rather unclear. We hypothesize that in the central nervous system (CNS) endogenous mechanisms of resilience or regeneration exist that critically contribute to the outcome of the inflammatory attack. Despite intensive research, mostly based on the experimental autoimmune encephalomyelitis (EAE) mouse model, the mechanisms underlying neurodegeneration and -regeneration are still unclear. For our studies, we have chosen to develop a human in-vitro CNS-like culture system implying the differentiation of human neural stem cells (hNSCs) and induced pluripotent stem cells (iPSCs) to neurons and astrocytes to establish a cell culture based model for studying inflammation-related neuronal damage and regeneration.
We have established protocols for a rapid and robust differentiation of hNSCs into neurons and astrocytes from H9-derived embryonal hNSCs or iPSCs from MS patients with a benign or accumulating disease course. Additionally, we study a Neurogenin (ngn)-2 transgenic iPSC culture, which drives the differentiation of homogenous glutamatergic neurons. The generated neuron- and astrocyte-like cells were characterized by immunofluorescence and qPCR using typical neuronal (MAP2, Tubb3, TH, CHAT, SLC6A4, and vGlut) and astrocytic markers (GFAP, S100beta, AQP4, EAAT1, ALDH1L1). For studying inflammation-related damage formation in neurons and neuron-/astrocyte- co-cultures, we investigated cell death and stress responses by the detection of structural changes in betaIII-tubulin and cleavage of caspase-3. The functional activity of neuron and astrocyte cultures were determined by Ca2+ imaging. We identified cytokine-specific neuronal network changes, highlighted by altered betaIII-tubulin expression, as well as glutamate induced effects in our culture model.
To conclude, we established an in-vitro CNS-like culture model, which is suitable to study aspects of neurodegeneration, as well as excitotoxicity. This may serve as a platform to explore potential mechanisms of neurodegeneration and resilience in well-characterized patient cohorts and might be helpful for the development of new treatment strategies.
Disclosure: Marlen Alisch, Janis Kerkering and Lil Meyer-Arndt contributed equally to this work. The authors report that there are no conflicts of interest. This work was funded by Gemeinnützige Hertie Stiftung.

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