5 years ago

Neural differentiation of bone marrow mesenchymal stem cells with human brain-derived neurotrophic factor gene-modified in functionalized self-assembling peptide hydrogel in vitro

Weihong Xu, Hongbin Luo, Xuping Lin, Huifen Zhong, Nanwen Zhang, Changsheng Wang, Yunda Hong, Lin Yang, Changsheng XU, Guisheng Liu, Xiaokun Chen, Zhiwei Liu, Xinyi Cai
Objective: To investigate the biocompatibility and differentiation of human brain-derived neurotrophic factor (hBDNF) gene-modified bone marrow mesenchymal stem cells (hBDNF-rMSCs) in a functionalized self-assembling peptide hydrogel. Methods: hBDNF were engineered in rMSCs using adenovirus vector and the enhanced green fluorescence protein (eGFP) was used as a reporter gene. MSC-specific surface markers (CD90,CD29 and CD45)were used for identifying rMSCs. Fluorescence microscope was used to detect the transfection of rMSCs. hBDNF-rMSCs and control cells (eGFP-rMSCs) were seeded in a functional self-assembling peptide hydrogel (RADA16-PRG hydrogel) and a control hydrogel (RADA16 hydrogel). Cells were divided into three groups (hBDNF-rMSCs + RADA16 Hydrogel, hBDNF-rMSCs + RADA16-PRG Hydrogel and eGFP-rMSCs+ RADA16-PRG Hydrogel) and a control group (eGFP-rMSCs + RADA16 Hydrogel). Cell growth, cell proliferation, expression of hBDNF-mRNA, the level of hBDNF, neuron-specific enolase (NSE), and glial fibrillary acidic protein (GFAP) protein were analyzed for each group. Result: rMSCs were positive for CD90 and CD29 and negative for CD45, green fluorescence was strongly visible at 72h after transfection. Compared with control group,the expression of hBDNF-mRNA and levels of hBDNF protein in both hBDNF group were significantly increased (P < 0.01),the cell growth, cell proliferation and levels of NSE and GFAP protein were significantly increased in three groups (P < 0.01). Cell growth, cell proliferation,expression of hBDNF-mRNA and levels of hBDNF,NSE and GFAP protein in hBDNF-rMSCs + RADA16-PRG Hydrogel Group were significantly higher than that of hBDNF-rMSCs + RADA16 Hydrogel Group (P < 0.01). Conclusion: Bone marrow mesenchymal stem cells can be induced into neural cells by the human brain-derived neurotrophic factor gene in a RADA16-PRG functionalized self-assembling peptide hydrogel. This article is protected by copyright. All rights reserved

Publisher URL: http://onlinelibrary.wiley.com/resolve/doi

DOI: 10.1002/jcb.26408

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