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

J Cell Biochem. 2019 Mar;120(3):2828-2835. doi: 10.1002/jcb.26408. Epub 2018 Dec 9.

Abstract

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 was engineered in rMSCs using adenovirus vector and the enhanced green fluorescence protein (eGFP) was used as a reporter gene. Mesenchymal stem cell-specific surface markers (CD90, CD29, and CD45) were used for identifying rat-derived MSCs. 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.

Results: rMSCs were positive for CD90 and CD29 and negative for CD45, green fluorescence was strongly visible at 72 hours 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 MSCs can be induced into neural cells by the human brain-derived neurotrophic factor gene in a RADA16-PRG functionalized self-assembling peptide hydrogel.

Keywords: bone marrow mesenchymal stem cells; differentiation; human brain-derived neurotrophic factor gene; hydrogel; self-assembling peptide.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain-Derived Neurotrophic Factor / genetics*
  • Brain-Derived Neurotrophic Factor / metabolism
  • Cell Differentiation*
  • Cell Proliferation
  • Cell Shape
  • Humans
  • Hydrogels / chemistry*
  • Mesenchymal Stem Cells / cytology*
  • Neurons / cytology*
  • Peptides / chemistry*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Rats, Sprague-Dawley

Substances

  • Brain-Derived Neurotrophic Factor
  • Hydrogels
  • Peptides
  • RNA, Messenger
  • BDNF protein, human