Brain-derived neurotrophic factor released from engineered mesenchymal stem cells attenuates glutamate- and hydrogen peroxide-mediated death of staurosporine-differentiated RGC-5 cells

详细信息    查看全文

• 作者：Matthew M. Harper ; Laura Adamson ; Bas Blits ; Mary Bartlett Bunge ; Sinisa D. Grozdanic ; Donald S. Sakaguchi
• 关键词：neuroprotection ; mesenchymal stem cells ; BDNF ; glaucoma ; ganglion cells ; RGC-5
• 刊名：Experimental Eye Research
• 出版年：2009
• 出版时间：October 2009
• 年：2009
• 卷：89
• 期：4
• 页码：538-548
• 全文大小：869 K

文摘

The purpose of this study was to determine the viability of cell-based delivery of brain-derived neurotrophic factor (BDNF) from genetically modified mesenchymal stem cells (MSCs) for neuroprotection of RGC-5 cells. RGC-5 cells were differentiated with the protein kinase inhibitor staurosporine (SS) and exposed to the cellular stressors glutamate or H₂O₂. As a neuroprotective strategy, these cells were then co-cultured across a membrane insert with mesenchymal stem cells (MSCs) engineered with a lentiviral vector for production of BDNF (BDNF-MSCs). As a positive control, recombinant human BDNF (rhBDNF) was added to stressed RGC-5 cells. After SS-differentiation RGC-5s developed neuronal-like morphologies, and a significant increase in the proportion of RGC-5s immunoreactive for TuJ-1 and Brn3a was observed. Differentiated RGC-5s also had prominent TrkB staining, demonstrating expression of the high-affinity BDNF receptor. Treatment of SS-differentiated RGC-5s with glutamate or H₂O₂, produced significant cell death (56.0 ± 7.02 and 48.90 ± 4.58 % of control cells, respectively) compared to carrier-solution treated cells. BDNF-delivery from MSCs preserved more RGC-5 cells after treatment with glutamate (80.0 ± 5.40 % cells remaining) than control GFP expressing MSCs (GFP-MSCs, 57.29 ± 1.89 % , p < 0.01). BDNF-MSCs also protected more RGC-5s after treatment with H₂O₂ (65.6 ± 3.47 % ) than GFP-MSCs (46.0 ± 4.20 % , p < 0.01). We have shown survival of differentiated RGC-5s is reduced by the cellular stressors glutamate and H₂O₂. Additionally, our results demonstrate that genetically modified BDNF-producing MSCs can enhance survival of stressed RGC-5 cells and therefore, may be effective vehicles to deliver BDNF to retinal ganglion cells affected by disease.