PhD Scientific Days 2024

Poster Session E - Molecular Medicine 2.

Transdifferentiation of human dental pulp-derived mesenchymal stem cells into neurosphere forming neurons and transplantation into aganglionic hindgut

Text of the abstract

Introduction: The enteric nervous system (ENS) regulates the function of the gastrointestinal tract. Numerous neurodevelopmental disorders underlie its defective function, mostly affecting the differentiation of neural stem cells of neural crest origin. Treatments for these diseases are limited, no disease-modifying surgery, treat the symptoms only, and no stem cell therapy available yet. Transdifferentiation, whereby patient-derived somatic cells are reprogrammed into neuronal lineage without going through an intermediate proliferative pluripotent stem cell stage, provides an alternative strategy for personalized stem cell therapy to treat congenital neurointestinal diseases.

Aims: Human dental pulp-derived mesenchymal stem cells (DPSCs), similar to the ENS, are neural crest-derived cells found in adult teeth. Our primary aim is to use DPSCs for intestinal transplantation to restore nervous tissue in neurocristopathies affecting the ENS.

Methods: We initiated our experimental work by generating neural cell aggregates (neurospheres) from DPSCs, directly reprogrammed toward a neural fate using viral transfection. Immunocytochemical methods were utilized to characterize the neurospheres' phenotype, and their functionality was tracked by transplanting them into aganglionic segments of 5-day-old chicken embryonic ceca and proximal hindgut.

Results: Serial sections through the induced neurospheres generated from human DPSC neurospheres demonstrate ubiquitous TUJ1+ neuronal expression and scattered SOX10+ precursors. The neurospheres were transplanted into the cecal region of E5 chick embryos and cultured on the chorioallantoic membrane of host E9 chick embryos or cultured in 3D collagen gel for 72 hours. After the grafts were harvested serial sections were stained with antibodies that specifically recognize human-derived cells and neurons.

Conclusion: Our findings suggest that neurospheres generated from DPSCs hold significant potential as a therapeutic avenue for neurointestinal disorders, but only in early stages of neural differentiation. We need to further investigate to enhance the cells' migration capabilities.

Funding: Supported by the ÚNKP-23-3-II, OTKA-K-138664, Semmelweis 250+ Scholarship for PhD Excellence