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


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.
Department of Pediatric Surgery, Medical Faculty Mannheim, University of Heidelberg, Theodor-Kutzer-Ufer 1-3, 68167 Mannheim, Germany.
Cornelia Irene
CI
Hagl
Ulrich
U
Rauch
Markus
M
Klotz
Sabine
S
Heumüller
David
D
Grundmann
Sabrina
S
Ehnert
Ulrike
U
Subotic
Stefan
S
Holland-Cunz
Karl-Herbert
KH
Schäfer
D002449:Cell Aggregation; D002454:Cell Differentiation; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D060833:Cellular Microenvironment; D002675:Child, Preschool; D004797:Enzyme-Linked Immunosorbent Assay; D005727:Ganglia, Spinal / Q000378:metabolism / Q000473:pathology; D041981:Gastrointestinal Tract / Q000473:pathology*; D005786:Gene Expression Regulation; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000235:genetics / Q000378:metabolism; D006627:Hirschsprung Disease / Q000235:genetics / Q000473:pathology* / Q000601:surgery; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D009124:Muscle Proteins / Q000378:metabolism; D009197:Myenteric Plexus / Q000254:growth & development* / Q000473:pathology*; D016501:Neurites / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014020:Tissue Extracts; D016212:Transforming Growth Factor beta / Q000378:metabolism
D051100:Glial Cell Line-Derived Neurotrophic Factor; D009124:Muscle Proteins; D012333:RNA, Messenger; D014020:Tissue Extracts; D016212:Transforming Growth Factor beta