Isolation and cultivation of neuronal precursor cells from the developing human enteric nervous system as a tool for cell therapy in dysganglionosis

Rauch, Ulrich; Hänsgen, Andrea; Hagl, Cornelia Irene; Holland‐Cunz, Stefan; Schäfer, Karl‐Herbert

doi:10.1007/s00384-005-0051-z

Cited by 100 publications

(89 citation statements)

References 19 publications

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“…Human enteric precursors have become a powerful tool for studying early embryonic stages of gut development. [25][26][27][28] In this study, we have evaluated the differences in the expression patterns of pluripotency stem cell marker genes between precursors derived from HSCR-NLBs and controlNLBs. We have obtained different expression levels in a group of genes (NESTIN, FN1, LAMC1, PECAM, SEMA3A, PAX6, and DNMT3B) between the two types of samples.…”

Section: Discussionmentioning

confidence: 99%

“…It is of interest to note that the three mutations are located in a region of the protein that interacts with DNMT1 to perform the establishment of a de novo methylation pattern that occurs during embryonic development. 25 Therefore, these mutations may hamper the appropriate methylation patterns and the regulation of gene expression required for proper embryonic development. The failure in the regulation of gene expression during the development of the ENS could be one of the mechanisms triggering HSCR in these patients.…”

Section: Discussionmentioning

confidence: 99%

“…These NLBs have been used as a source of enteric precursor cells that can be transplanted to restore the contractile properties of aganglionic bowel by neurally mediated mechanisms. [25][26][27][28] In the present study, we have used such NLBs to search for new genes that could be implicated in ENS formation and in the pathogenesis of HSCR. Given that neural crest pluripotency is essential for proper ENS development in particular, we have studied the expression profiles of pluripotency-associated genes in enteric neural precursors.…”

Section: Methodsmentioning

confidence: 99%

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Involvement of DNMT3B in the pathogenesis of Hirschsprung disease and its possible role as a regulator of neurogenesis in the human enteric nervous system

Torroglosa

Enguix-Riego

Fernández

et al. 2014

Genetics in Medicine

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Involvement of DNMT3B in the pathogenesis of Hirschsprung disease and its possible role as a regulator of neurogenesis in the human enteric nervous system

Torroglosa

Enguix-Riego

Fernández

et al. 2014

Genetics in Medicine

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“…In this study, we established a stable and convenient method to isolate and culture ENSCs on the basis of previous studies (Le et al 2004;Farlie et al 2004;Ulrich Rauch et al 2006). We found out that the numbers of ENSCs obtained from the adult and new born mice were very low.…”

Section: Discussionmentioning

confidence: 99%

Prospective identification and culture of rat enteric neural stem cells (ENSCs)

et al. 2014

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Hirschprung's disease (HD), a very common congenital abnormality in children, occurs mainly due to the congenital developmental defect of the enteric nervous system. The absence of enteric ganglia from the distal gut due to deletion in gut colonization by neural crest progenitor cells may lead to HD. The capacity to identify and isolate the enteric neuronal precursor cells from developing and mature tissues would enable the development of cell replacement therapies for HD. However, a mature method to culture these cells is a challenge. The present study aimed to propose a method to culture enteric neural stem cells (ENSCs) from the DsRed transgenic fetal rat gut. The culture medium used contained 15 % chicken embryo extract, basic fibroblast growth factor, and epidermal growth factor. ENSCs were cultured from embryonic day 18 in DsRed transgenic rat. Under inverted microscope and fluorescence staining, ENSCs proliferated to form small cell clusters on the second day of culture. The neurospheres-like structure were suspended in the medium, and there were some filaments between the adherent cells from day 3 to day 6 of the culture. The neurospheres were formed by ENSCs on day 8 of the culture. Network-like connections were formed between the adherent cells and differentiated cells after adding 10 % FBS. The differentiated cells were positive for neurofilament and glial fibrillary acidic protein antibodies. The present study established a method to isolate and culture ENSCs from E18 DsRed transgenic rats in the terminal stage of embryonic development. This study would offer a way to obtain plenty of cells for the future research on the transplantation of HD.

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“…Recent studies however, have provided evidence that a population of neural crest-derived stem cells exists after birth, in the postnatal and adult rodent (Bondurand et al 2003;Kruger et al 2002) and human (Rauch et al 2006a) (Almond et al 2007) gut. To date, several different methods have been used to identify and select these cells from postnatal and adult rodent intestine.…”

Section: Introductionmentioning

confidence: 99%

Neural progenitors from isolated postnatal rat myenteric ganglia: Expansion as neurospheres and differentiation in vitro

et al. 2008

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Identification of the stem cell niche is crucial for understanding the factors that regulate these cells. Rodent enteric neural crest-derived stem cells have previously been isolated by flow cytometry and culture of cell suspensions from the outer smooth muscle layers or the entire gut wall from postnatal and adult animals. Such cell suspensions contain a mixture of cell types, including smooth muscle, fibroblasts and cells associated with the vasculature and extrinsic innervation. Thus these preparations may be contaminated by stem cells associated with extrinsic sensory and autonomic nerves and by other types of stem cell that reside in the gut. Here we describe a different approach, similar to that recently used for infant human gut, to obtain enteric ganglionderived cells, with properties of neural progenitor cells, using isolated myenteric ganglia from postnatal rat ileum. Myenteric ganglia were separated from the gut wall, dispersed and resulting cell dissociates were plated in non-adherent culture conditions with EGF and FGF-2. Under these conditions neurospherelike bodies (NLB) developed. Cells in NLB incorporated BrdU and expressed the stem cell marker nestin but not the pan-neuronal marker PGP 9.5. Upon growth factor withdrawal some BrdU-immunopositive cells assumed the morphology of neurons and expressed PGP 9.5; others were flattened and expressed the glial cell marker GFAP. This work therefore provides evidence that neural crest-derived progenitors in the postnatal rat gut are located in the myenteric plexus, and shows that these cells can be expanded and differentiated in NLB in vitro.

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Isolation and cultivation of neuronal precursor cells from the developing human enteric nervous system as a tool for cell therapy in dysganglionosis

Abstract: The first results using isolated enteric neurospheres in aganglionic bowel are quite promising and are a basis to develop an appropriate cell therapy for all kinds of dysganglionosis, especially for cases where a surgical approach is not sufficient or not even possible.

Cited by 100 publications

References 19 publications

Involvement of DNMT3B in the pathogenesis of Hirschsprung disease and its possible role as a regulator of neurogenesis in the human enteric nervous system

Involvement of DNMT3B in the pathogenesis of Hirschsprung disease and its possible role as a regulator of neurogenesis in the human enteric nervous system

Prospective identification and culture of rat enteric neural stem cells (ENSCs)

Neural progenitors from isolated postnatal rat myenteric ganglia: Expansion as neurospheres and differentiation in vitro

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