Transplantation of PSA-NCAM expressing embryonic stem cell-derived neural progenitors in organotypic mouse gut cultures

Defective migration of neural precursors in the developing gut leads to intestinal aganglionosis (Hirschsprung's disease). To overcome limits associated with the current surgical treatment of this common birth disorder we are exploring regenerative tissue repair of the gut using ES cell-derived neural cells.

During embryonic development, the enteric nervous system (ENS) is derived from neural crest cells. Bone morphogenetic proteins (BMPs) are known to regulate important aspects of ENS progenitor differentiation and migration.

PSA-NCAM is regarded as a key modulator of neural migration. Recently, BMPs have been shown to exhibit neural crest-inducing properties in embryonic stem cell-derived neural precursors. Immature BMP-induced peripheral neurons exhibit strong PSA-NCAM expression.

Exploiting these observations, we combined PSA-NCAM immunopanning and BMP treatment to enriche populations of peripheral neuronal precursors. Upon in vitro differentiation these progenitors segregate into GABAergic, glutamatergic, cholinergic and calretinin-positive phenotypes. After in vitro transplantation they incorporate into organotypic cultures of the mouse gut, where they predominantly aquire a cholinergic, Ret-positive fate. These data indicate that ES cell-derived peripheral neural precursors might serve as a donor source for enteric nervous system repair.