NEUROGENESIS FROM AGANAGLIONIC ENTERIC TISSUE

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Excerpt

Intestinal prenatal neurogenesis is due to neural crest cells (NCC). Three different research groups have identified the persistence of neural stem cells (NSC) in the postnatal intestine. At least some of the postnatal intestinal NSC are believed to be of neural crest origin. Reported differences in culture techniques and NSC behaviour suggest there may be more than one NSC type. To explore this, we studied aganglionic intestinal tissue biopsies from 5 Hirschsprung's disease (HD) patients, where NCC are absent and attempted to grow in culture, newly differentiated neurons. For this we used a mechanichal and enzymatic cellular dissociation technique with which we have accomplished in vitro neurogenesis using normal postnatal intestine (Suarez-Rodriguez and Belkind-Gerson, Stem Cells, Dec 2004). After a week, all biopsies produced small colonies of neuron-like cells positive for tau, synaptophysin, MAP2 and neurofilaments, extending long cellular processes and forming intercellular connections. Electrophysiologic (patch-clamp) studies were performed on neuron-like cells from two of the patients and revealed that action potentials could be elicited. This study shows that in vitro, from HD aganglionic tissue, it is possible to obtain neuron-like cells morphologically, functionally and expressing several neuronal-specific proteins. These results suggest that the postnatal intestine, in addition to neural crest-derived stem cells may posses another cellular source for neurogenesis.
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