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

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.