AN<EM> IN VITRO</EM> GLIAL SCAR MODEL TO ASSESS SURVIVAL AND INTERACTIONS OF SCHWANN-LIKE CELLS WITH NEURITES FOR NEURAL REGENERATION

CNS regeneration remains a significant challenge in regenerative medicine. Cell therapy, particularly using Schwann-like phenotypes such as olfactory ensheathing cells (OEC), offers a promising solution due to their pro-regenerative molecular and functional characteristics. To overcome the limitations of obtaining and maintaining OECs, the differentiation of bone marrow mesenchymal stem cells (BM-MSC) into a Schwann-like phenotype (BM-MSCdiff) has been proposed. This study aimed to observe the phenotype, survival, and cellular interactions of OECs, BM-MSCs, and BM-MSCdiff with neurites in an in vitro glial scar environment. An in vitro glial scar model was established using primary cortical neuron cultures followed by a scratch assay. OEC, BM-MSC, and BM-MSCdiff were seeded separately 3h post-injury. After 72 hours, immunocytochemistry was performed using anti-p75, anti-NF-L, DAPI to evaluate interactions; cell phenotype was evaluated separately with anti-p75, anti-GFAP and anti-CD90. All cell types successfully established in the model. OECs displayed spindle-like morphology, and positioned near the injury border and co-localized with p75 and NF-L, indicating direct interaction with neurites. BM-MSCs filled the injury gap with a characteristic fibroblast-like morphology but showed no co-localization or neurite interaction. BM-MSCdiff localized near the injury border and center, exhibiting spindle-like morphology but no co-localization, representing a mixed interaction profile. The results demonstrate that this glial scar model shows promising characteristics for neural regeneration analysis. While all cell types proved capable of establishment, further research is required to evaluate the specific interactions of BM-MSCdiff and their functional role in neuronal regeneration.