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Activated adult microglia influence retinal progenitor cell proliferation and differentiation toward recoverin-expressing neuron-like cells in a co-culture model

  • Basic Science
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Abstract

Purpose

Microglia contribute to immune homeostasis of the retina, and thus act as a potential regulator determining successful repair or retinal stem cell transplantation. We investigated the interaction between human microglia and retinal progenitor cells in cell co-culture to further our exploration on developing a new therapeutic strategy for retinal degeneration.

Methods

Microglia and retinal progenitor cultures were developed using CD11b+ and CD133+, respectively, from adult donor retina. Microglia activation was developed using interferon-gamma and lipopolysaccharide. Retinal progenitor differentiation was analysed in co-culture with or without microglial activation. Retinal progenitor proliferation was analysed in presence of conditioned medium from activated microglia. Phenotype and function of adult human retinal cell cultures were examined using cell morphology, immunohistochemistry and real-time PCR.

Results

By morphology, neuron-like cells generated in co-culture expressed photoreceptor marker recoverin. Neurospheres derived from retinal progenitor cells showed reduced growth in the presence of conditioned medium from activated microglia. Delayed retinal progenitor cell migration and reduced cellular differentiation was observed in co-cultures with activated microglia. In independent experiments, activated microglia showed enhanced mRNA expression of CXCL10, IL-27, IL-6, and TNF-alpha compared to controls.

Conclusion

Adult human retina retains retinal progenitors or potential to reprogram cells to then proliferate and differentiate into neuron-like cells in vitro. Human microglia support retinal progenitor differentiation into neuron-like cells, but such capacity is altered following microglial activation. Modulating microglia activity is a potential approach to promote retinal repair and facilitate success of stem-cell transplantation.

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Acknowledgments

We thank Dr. Lindsay B. Nicholson for advice on data analysis and manuscript writing. The work was supported through Guide Dogs for the Blind Association, UK (OR2009-02e). A.D.D. is partially supported through funding. This work was partly supported by the National Institute for Health Research (NIHR) Biomedical Research Centre based at Moorfields Eye Hospital NHS Foundation Trust and UCL Institute of Ophthalmology. The views expressed are those of the author (s) (A.D.D.) and not necessarily those of the NHS, the NIHR, or the Department of Health.

Disclosure of potential conflicts of interest

The authors indicate no potential conflicts of interest.

Author information

Authors and Affiliations

  1. School of Clinical Sciences, University of Bristol, Bristol, United Kingdom

    Yunhe Xu, David A. Copland, Jian Liu, M. John Armitage & Andrew D. Dick

  2. Bristol Eye Hospital, Lower Maudlin Street, Bristol, United Kingdom

    Balini Balasubramaniam

  3. Corneal Transplant Service Bristol Eye Bank, Bristol, United Kingdom

    M. John Armitage

  4. National Institute for Health Research Biomedical Research Centre at Moorfields Eye Hospital NHS Foundation Trust and UCL Institute of Ophthalmology, University Hospitals Bristol NHS, Foundation Trust, and University of Bristol, Bristol, United Kingdom

    Andrew D. Dick

  5. University of Bristol, Level 2, Learning and Research Building, Southmead Hospital, Westbury-on-Trym, Bristol, BS10 5NB, United Kingdom

    Yunhe Xu

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  1. Yunhe Xu
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  2. Balini Balasubramaniam
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Correspondence to Yunhe Xu.

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Supplement Fig. 1

Co-expression of Iba-1 in CD11b+ cells by immunofluorescence. CD11b+ cells isolated from donor eye were further cultured for 10 days. Cells were then fixed in paraformaldehyde (4 %), permeabilized in Triton-100, and then stained using mouse-anti-human mAb, Iba-1 (Abcam; 1:100) overnight at 4 °C, and detected following 2 h incubation with FITC-labelled bovine anti-mouse IgG (Santa Cruz; 1:200) at room temperature, mounted and counterstained with DAPI (Vector Laboratories). Photos show phase image (a), DAPI image (b), FITC image (c) and merged image (d); scale bar = 50 μm (DOCX 2195 kb)

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Xu, Y., Balasubramaniam, B., Copland, D.A. et al. Activated adult microglia influence retinal progenitor cell proliferation and differentiation toward recoverin-expressing neuron-like cells in a co-culture model. Graefes Arch Clin Exp Ophthalmol 253, 1085–1096 (2015). https://doi.org/10.1007/s00417-015-2961-y

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  • DOI: https://doi.org/10.1007/s00417-015-2961-y

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