Oligodendrojenez modellemesi için 3 Boyutlu (3B) çoklu ve mikropaternli GelMA Platformu

The occurrence of disturbances in myelination can lead to the development of a range of disorders. A reliable and biologically mimicking in vitro model of oligodendrogenesis is of necessity to aid in elucidating the pathogenesis of demyelinating diseases. Oligodendrogenesis and myelination are highly dependent on the function of extracellular matrix (ECM) components. The recent advancements in biomaterials illustrated that hydrogels have tremendous potential to mimic the ECM of central nervous system (CNS). It remains elusive whether the characteristics of hydrogels can manage to regulate the differentiation and maturation of oligodendrocyte (OL) lineage cells and provide a suitable model without the need for primary cells or in vivo models. Here, this study examines the impact of a multiplexed and micropatterned 3 Dimensional (3D) Gelatin Methacrylate (GelMA) platform on the viability, and spheroid formation of Human Oligodendroglioma (HOG) cells under different UV exposure conditions, while also exploring the maturation status. Both proliferation and spheroid formation were enhanced with prolonged UV exposure durations, illustrating the importance of ECM dynamics on cellular behaviour. The cell viability on day 5 was the greatest at 50 seconds of exposure condition. The formation of spheroids was discernibly observed on day 5 at exposure conditions of 40 and 50 seconds, with the latter showing a notable effect. Maintenance in GelMA induced the mRNA expression of transcription factors OLIG2, SOX2 and SOX10; OPC markers PDGFRα and NG2; and OL markers CNPase, MBP, and MOG. The expression of GFAP was not detected in the case of 3D whereas it was found to be traceably expressed in 2D cultured cells, suggesting a possible rescue from astrocyte lineage in 3D. Immunofluorescence microscopy confirmed the expression of MBP and MOG at the protein level. These results imply the regulatory effect of the GelMA environment on the expression profile of HOG cells, altering their fate towards OL lineage. To ascertain whether alterations in the expression of OL genes are exclusive to 3D maintenance, a separate treatment of UV and GelMA was applied to HOG cells cultured in a 2D environment. Neither UV treatment nor GelMA addition made significant changes on the expression profile of HOG cells. In conclusion, this platform appears to be a noteworthy culture model for oligodendrogenesis, warranting further investigation. Coculture of SH-SY5Y cells with HOG cells was conducted to see if any interaction would occur. Although SH-SY5Y cells were not differentiated into mature neurons, their interaction with HOG cells was evident, leading to accumulation near HOG spheroids with the latter crosslinking exhibiting a notable effect. Further investigation with the usage of mature neurons and oligodendrocyte lineage cells within the hydrogel may aid in enlightening whether the platform also serve as a model for myelination. Key words: Oligodendrocytes, oligodendrogenesis, 3D-hydrogel Model, GelMA, myelination