Cultivation and Neural Differentiation of Embryonic Cerebrospinal Fluid Treated Adipose Stem Cells on the Scaffold of Amniotic Membrane

Adipose stem cells (ASCs) are ideal candidates for cell therapy of neurological disorders. In vitromethods require the use of a variety of growth factors and multi-step protocols to induce neuronal differentiation. This study was aimed to assess the neural differentiation of adipose stem cells in a co-culture system.

ASCs were obtained from male Wistar rats and were characterized, using flow cytometry. Harvested ASCs were cultured on a scaffold prepared from amniotic membrane (AM). Cerebrospinal fluid (CSF) was collected from rat embryos and was added toculture medium for 7 days. Structure of scaffold and cell attachment was assessed through scanning electron microscopy (SEM). Neural differentiation of ASCs in the co-culture system was confirmed with immunofluorescence (IF) staining for β-tubulin III andMAP-2 markers.

SEM results confirmed the decellularization of AM and attachment of ASCs on the AM derived scaffold. MTT assay revealed that ASCs proliferated on AM significantly during the 7 days of culture. IF data confirmed that the CSF treated cells were expressed by β-tubulin III and MAP-2 but untreated cells were negative for the expression of neural markers.

Cultivation of ASCs on the scaffold and their treatment with CSF induced them into the neural lineage fate in the absenceof any chemical inducing factor. This method of co-culture may represent a new method to improve in vitroneural differentiation of ASCs.