In vitro study of neural stem cells and activated Schwann cells cocultured on electrospinning polycaprolactone scaffolds

Abstract

Baoyou Fan,1,2,* Xianhu Zhou,1,2,* Lina Wang,3,* Zhijian Wei,1,2 Wei Lin,1,2 Yiming Ren,1,2 Guidong Shi,1,2 Xin Cheng,1,2 Lianyong Wang,3 Shiqing Feng,1,2 1International Science and Technology Cooperation Base of Spinal Cord Injury, Department of Orthopedic Surgery, Tianjin Medical University General Hospital, 2Tianjin Neurological Institute, Key Laboratory of Post-neuroinjury Neuro-repair and Regeneration in Central Nervous System, Ministry of Education and Tianjin City, 3Key Laboratory of Bioactive Materials, Ministry of Education, College of Life Sciences, Nankai University, Tianjin, People’s Republic of China *These authors contributed equally to this work Background: To investigate the biocompatibility of electrospinning polycaprolactone (PCL) fiber scaffolds and coculture system, which consisted of neural stem cells (NSCs) and activated Schwann cells (ASCs). Materials and methods: ASCs were isolated from sciatic nerves, ligated for 7 days, in 4-week-old Wistar rats, and the NSCs were isolated from the hippocampus of E14.5 Wistar rat embryos. ASCs, NSCs and ASCs combined with NSCs were 3D cultured on the electrospinning PCL fiber scaffolds. Crystal violet staining was used to find the suitable density of ASCs for growth, and the proliferation of NSCs and ASCs were tested by Cell Counting Kit (CCK)-8 assay, and cell adhesion, differentiation of NSCs and myelin basic protein (MBP) expression of ASCs were observed by laser confocal microscopy. Distribution and morphology were assessed by scanning electron microscopy. Results: The average diameter of fibers in electrospinning PCL scaffolds was approximately 7.93±1.41 μm. ASCs could grow well at the density of 2×104/cm2, and a certain number of cells extended along the longitudinal axis of fibers, and the shape of the cells was spindle, which was consistent with crystal violet staining results. The CCK-8 experiment showed ASCs could proliferate gradually on the PCL scaffold within 7 days, as well as NSCs, and NSCs differentiated into astrocytes, neurons and oligodendrocytes on the PCL scaffold; PCL scaffolds could improve the differentiation rate of neurons. After NSCs and ASCs were cocultured on electrospinning PCL scaffolds, ASCs could express MBP and NSCs could differentiate into neurons, which distributed around those ASCs expressing MBP.Conclusion: Electrospinning PCL fibrous scaffolds showed good biocompatibility, and the fibers had an inducing effect on the distribution of ASCs. NSCs and ASCs cultured on electrospinning PCL scaffolds could form 3D culture system, and NSCs could differentiate into neurons which distributed around the ASCs expressing MBP. Keywords: neural stem cells, Schwann cells, PCL, scaffold, electrospinning 

Keywords

• neural stem cells
• Schwann cells
• PCL
• scaffold