The microenvironment in the Hirschsprung's disease gut supports myenteric plexus growth
The microenvironment in the Hirschsprung's disease gut supports myenteric plexus growth
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DOI：
10.1007/s00384-012-1411-0
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发表时间：
2012-06-01
影响因子：
2.8
通讯作者：
Schaefer, Karl-Herbert
中科院分区：
文献类型：
--
作者：
Hagl, Cornelia Irene;Rauch, Ulrich;Schaefer, Karl-Herbert
神经嵴源性干细胞（NCSC）移植是治疗先天性巨结肠症（HSCR）的有效替代方法。待移植的细胞应该找到合适的微环境来存活和分化。本研究采用离体培养的方法，研究了HSCR平滑肌蛋白提取物对离体肠肌间神经丛细胞、离体背根神经节和NCSC的影响，探讨了这种微环境效应的质量。解剖平滑肌并均质化。采用ELISA法测定各节段匀浆中胶质细胞源性神经营养因子（GDNF）和转化生长因子β-1（TGF β-1）的浓度。肌间神经丛和分离的背根神经节（DRG）的文化，以及NCSC暴露于蛋白质提取物来自神经节和无神经节HSCR节段，其影响后神经突生长，生存，和分支进行了评估。的量的因素之间的个别段，也从病人到病人有很大的变化。可检测到四种主要的表达模式。虽然与对照组相比，所有供试浸提液均导致神经突生长显著增加，但近端节段浸提液往往具有更显著的影响。在一个实验中，测试了来自单个患者的所有单个节段的提取物。神经突生长，神经元的生存，和分支模式不同的节段，但所有的HSCR肌肉蛋白提取物增加神经元的生存和网络的形成。来自无神经节肠的平滑肌蛋白支持肌间神经元和NCSC的存活和生长，因此是神经干细胞治疗的合适靶点。
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.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-beta-1 (TGF beta-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.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.