PhD Scientific Days 2025

Theoretical and Translational Medicine I.

Experimental Modeling of Hirschsprung’s Disease and Neural Stem Cell Transplantation Using Embryo Manipulation Techniques

Name of the presenter

Jurenka Csenge Lili

Institute/workplace of the presenter

Department of Anatomy, Histology and Embryology

Authors

Csenge Lili Jurenka¹

1: Department of Anatomy, Histology and Embryology

Text of the abstract

Introduction: In vertebrate embryos, neural crest-derived stem cells from the cervical somite level migrate into the foregut mesenchyme and along the developing gut wall to form the enteric nervous system (ENS). If neural crest cell development or regulation is impaired, migration fails, leading to ENS underdevelopment and ganglion-free gut segments. This causes Hirschsprung’s disease, marked by distal colon aganglionosis and severe intestinal obstruction. Current treatment is surgical removal of the affected segment, often with complications. My research aimed to explore a new stem cell transplantation method in an experimental Hirschsprung’s disease model.

Aims and Methods: The first objective of my work was to develop an embryonic Hirschsprung’s disease (HD) model that presents distal colorectal aganglionosis without genetic modification and is suitable for modeling intestinal transplantation of neural stem cells through embryomanipulation experiments. For this, we performed both in vivo and in vitro experiments using 2-day-old chicken embryos injected with chondroitin sulfate proteoglycan (CSPG) (n=85), as well as neural tubes explanted from them. The ganglion-free hindguts, developing on the chorioallantoic membrane and treated with CSPG, were transplanted with cell aggregates (neurospheres) derived from embryonic quail gut wall neurons (n=8).

Results: In vitro, CSPG blocked early neural crest migration, unlike fibronectin, which promoted it. In vivo, CSPG injected into the cervical neural crest caused reduced hindgut innervation after 8 days of incubation. Transplanted neurospheres colonized the host tissue and formed enteric Hu+/Tuj1+ ganglia.

Conclusions: CSPG’s inhibitory effect on migration was confirmed in vitro and in vivo. Ganglion-free hindgut transplantation using neurospheres proved feasible and may support future ENS regeneration strategies.

Funding: OTKA-K-138664, EKÖP Scholarship