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Identifier

000434262

Title

Evaluation of NSC-based implants for a spinal cord injury mouse model

Alternative Title

Αξιολόγηση εμφυτευμάτων σε νευρικά βλαστικά κύτταρα, για μοντέλο ποντικού με τραύμα στο νωτιαίο μυελό

Author

Μπαμπούλα, Ευσταθία Ε.

Thesis advisor

Γραβάνης, Αχιλλέας

Reviewer

Χαραλαμπόπουλος, Ιωάννης
Τζεράνης, Δημήτριος

Abstract

Spinal Cord Injury (SCI) refers to a damage to the spinal cord or specific nerves located at the spinal canal’s end, which temporarily or permanently causes functional problems. SCI is recognized as a global health issue due to its complex and multifactorial pathophysiology and its significant impact on patient quality of life. In combination with failure of CNS neurons, to maintain their intrinsic growth capacity and the great inability to regenerate their axons following injury, SCI treatment strategies still remain controversial. NSPC-seeded scaffolds engrafted after the SCI are a promising SCI treatment, since they seem to be able to impact the lesion site and improve functional recovery. This study builds upon previous work of the IMBB Neural Tissue Engineering Lab on SCI grafts based on NSPCs-seeded porous collagen-based scaffolds (PCS). According to a recent study by Kouriantaki et al., NSPC-seeded PCS engrafted in a dorsal column crush SCI mouse model lead to significant improvement in locomotion recovery 10-12 weeks post injury. The objective of this thesis is to characterize the utilized NSPC-seeded grafts in more detail and evaluate the ability to modulate NSPCs fate once seeded in PCSs. NSPC fate characterization inside scaffolds utilized immunocytochemistry. Fluorescence microscopy images were utilized to quantify the percentage of cells that stain positive for markers of specific cell developmental stages and thereby discriminate different NSPCs subpopulations. Results demonstrate that in NSPC-seeded grafts the largest percentage of cells are NSPCs, characterized by the expression of Sox2, Nestin and GLAST, with the highest percentage being RGCs. However, results demonstrate that the cell population of such implants consisted also of GRPs and astrocytes (based on GFAP expression) and OPCs (based on PDGFRa expression). The lower percentage of Tuj1 positive cells, in comparison with the glial markers, indicates a preference of cell differentiation towards glial cells in PCSs. In comparison with 2D culture, where cells remain in the median stage of NSPCs, cells grown inside scaffolds culture seem to follow a stricter pattern, being either at the stage of NECs or even of more mature progenitors-GRPs. Inducing NSPCs towards a neural fate by RA treatment did not work in this system both in 3D culture (inside scaffolds) and in ordinary 2D monolayer culture. Even when treated with differentiation mediums, PCSs favor NSPC differentiation towards OPCs and enhanced the expression of NSPC markers over a longer time compared to ordinary 2D culture.

Language

English, Greek

Subject

Neural stem cells

Neuroimplants

Porous collagen scaffolds

Νευροεμφυτεύματα

Πορώδη ικριώματα κολλαγόνου

Τραύμα νωτιαίου μυελού

Issue date

2020-11-27

Collection