Comparison of the effect of stem cells derived from amniotic fluid and supernatant obtained from these stem cells on neurogenesis and reprogramming of astrocyte cells to neuroblast in spinal cord injury model

Abstract

Spinal cord injury (SCI) is damage to the spinal cord that resulted in irreversible neuronal loss, glial scar formation and axonal injury. Herein, we used the human amniotic fluid mesenchymal stem cells (hAF-MSCs) and their conditioned medium (CM), to investigate their ability in neuroblast and astrocyte production as well as functional recovery following SCI. Methods: Sixty-six adult rats were randomly divided into eleven groups (n=6), included: Control, SCI, SCI+CM-IP, SCI+DMEM-IP, SCI+CM-Focal, SCI+DMEM-Focal, SCI+MSCs, SCI + Astrocyte, SCI+Astrocyte+DMEM-IP, SCI + Astrocyte+ CM-IP and SCI + Astrocyte+ MSCs. Following laminectomy and SCI induction, DMEM, CM, MSCs, and Astrocytes were injected. Wester-blot was performed to explore the levels of the Sox2 protein in the MSCs-CM. The immunofluorescence staining against DCX and GFAP was done. Finally, Basso-Beattie-Brenham (BBB) locomotor test was conducted to assess the neurological outcomes. Results: Our results showed that the MSCs increased the number of endogenous DCX-positive cells and decreased the number of GFAP-positive cells. Also, MSCs and CM containing Sox2 factor can convert transplanted human astrocytes into neuroblasts, and the final results showed that the effect of MSCs is more effective than CM. Finally, fifteen days after the lesion, the improvement of motor function was evident in the treatment groups with CM and MSCs.