Proceedings of The Physiological Society

Europhysiology 2018 (London, UK) (2018) Proc Physiol Soc 41, PCB258

Poster Communications

The peripheral nervous system in vitro - Characterization of a co-culture system with human derived neurones and Schwann cells

C. Trinler1, C. Rösseler1, P. Hautvast1, A. Lampert1

1. Institute of physiology, RWTH Aachen, Aachen, Germany.

Peripheral neuropathy can be induced by inherited or acquired dysfunctions of the peripheral nervous system. The disease causing factors are largely unknown. Different cell types and cell-cell interactions, such as the axon-Schwann cell (SC) interaction, can be affected. In order to study these interactions in detail a reliable model is needed, which includes both neurones and SCs. Progress in the differentiation of human induced pluripotent stem cells (hiPSCs) into different cell types has led to efficient protocols generating human sensory neurones and SCs (Chambers et al., 2012; Kim et al., 2017). Previous approaches to establish a co-culture were achieved in a heterologous system of rodent and human cells (Clark et al., 2017) and with long-term protocols using human embryonic stem cells derived sensory neurones and SCs (Ziegler et al. 2011). By using a recently published protocol (Kim et al., 2017) we generate a homogeneous culture of hiPSCs derived SCs for co-culturing with hiPSCs derived sensory neurones, which we produce using a small molecule approach (Eberhardt et al., 2015, based on Chambers et al., 2012). The expression of typical SC markers, including S100, GFAP, p75 and Sox10, is approved by immunostaining and mRNA expression. We co-culture human SCs with iPSCs derived sensory neurones similar as described in Clark et al. 2017 for rodent SCs. Within an hour after adding SCs to iPSCs derived neurones, alignments of spindle-shaped SC-like cells to axonal structures are observed. To analyse the effect of SCs on sensory neurones in a co-culture system we study immunocytochemical characteristics and canonical markers in quantitative RT-PCR like MBP, Caspr and ion channel expression. In addition, we aim to study the electrophysiological aspects of nociceptors matured in the presence of SCs by using current clamp to measure action potential generation. This will help to explore whether SCs promote and accelerate the process of maturation of neurones. By providing a human disease model for neuropathy which more faithfully follows physiology, we aim to explore underlying disease mechanisms and potentially to support the development of new drugs.

Where applicable, experiments conform with Society ethical requirements