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The microenvironment in the Hirschsprung's disease gut supports myenteric plexus growth

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Abstract

Introduction

The transplantation of neural crest derived stem cells (NCSC) is a potent alternative for the treatment of Hirschsprung's disease (HSCR). Cells to be transplanted should find an appropriate microenvironment to survive and differentiate. Influences of HSCR-smooth-muscle-protein extracts upon isolated myenteric plexus cells, dissociated dorsal root ganglia and NCSC were studied in vitro to investigate the quality of this microenvironment effects.

Methods

Postnatal human gut from children undergoing colonic resection due to HSCR was divided in segments. Smooth muscle was dissected and homogenized. Glial-cell-line-derived-neurotrophic-factor (GDNF) and transforming-growth-factor-β-1 (TGFβ-1) concentration were measured in the homogenates from the individual segment using ELISA. Myenteric plexus and dissociated dorsal root ganglia (DRG) cultures, as well as NCSCs were exposed to protein extracts derived from ganglionic and aganglionic HSCR segments, and their effect upon neurite outgrowth, survival, and branching was evaluated.

Results and conclusions

The amount of the factors varied considerably between the individual segments and also from patient to patient. Four major expression patterns could be detected. While all extracts tested lead to a significant increase in neurite outgrowth compared to the control, extracts from proximal segments tended to have more prominent effects. In one experiment, extracts from all individual segments of a single patient were tested. Neurite outgrowth, neuronal survival, and branching pattern varied from segment to segment, but all HSCR-muscle-protein extracts increased neuronal survival and network formation. Smooth muscle protein from aganglionic bowel supports the survival and outgrowth of myenteric neurons and NCSCs and is so an appropriate target for neural stem cell treatment.

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Acknowledgements

We are grateful to Elvira Wink and Karin Kaiser for excellent technical support. The work was supported by the German Research Foundation (DFG SCHA 878-1 and 2).

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Authors and Affiliations

  1. Department of Pediatric Surgery, Medical Faculty Mannheim, University of Heidelberg, Theodor-Kutzer-Ufer 1-3, 68167, Mannheim, Germany

    Cornelia Irene Hagl, Sabine Heumüller & Karl-Herbert Schäfer

  2. Department of Computer Sciences and Microsystems Technology, University of Applied Sciences, Life Science, Amerikastraße 1, 66482, Zweibrücken, Germany

    Ulrich Rauch, Markus Klotz, David Grundmann & Karl-Herbert Schäfer

  3. Department of Traumatic Surgery, Technical University Munich, Ismaningerstraße 22, 81675, Munich, Germany

    Sabrina Ehnert

  4. Department of Pediatric Surgery, University of Zürich, Steinwiesstraße 75, 8032, Zürich, Switzerland

    Ulrike Subotic

  5. Department of Pediatric Surgery, University of Heidelberg, Im Neuenheimer Feld 110, 69120, Heidelberg, Germany

    Stefan Holland-Cunz

  6. Enteric Nervous System Working Group, Department of Pediatric Surgery Mannheim, University of Applied Sciences Kaiserslautern, Amerikastraße 1, 66482, Zweibrücken, Germany

    Karl-Herbert Schäfer

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  1. Cornelia Irene Hagl
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Correspondence to Karl-Herbert Schäfer.

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Hagl, C.I., Rauch, U., Klotz, M. et al. The microenvironment in the Hirschsprung's disease gut supports myenteric plexus growth. Int J Colorectal Dis 27, 817–829 (2012). https://doi.org/10.1007/s00384-012-1411-0

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