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

Introduction: Adipose stem cells (ASCs) are ideal candidates for cell therapy of neurological disorders. In vitro methods 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. Material and Methods: 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 to culture 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 and MAP-2 markers. Results: 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. Conclusion: Cultivation of ASCs on the scaffold and their treatment with CSF induced them into the neural lineage fate in the absence of any chemical inducing factor. This method of co-culture may represent a new method to improve in vitro neural differentiation of ASCs.