Abstract：
:Retinal inflammation is an integral component of many retinal diseases including diabetic retinopathy (DR), age-related macular degeneration (AMD) and retinopathy of prematurity (ROP). Inflammation is commonly initiated and perpetuated by myeloid-derived immune cells. In the retina, microglial cells are resident macrophages with myeloid origins, which acts as the first responders involved in the innate immune system. To understand the disease pathogenesis, the use of isolated retinal cell culture model is vital for the examination of multiple cellular responses to injury or trauma. The pig retina resembles human retina in terms of tissue architecture, vasculature, and topography. Additionally, it is a better model than the rodent retina because of the presence of the pseudomacula. In the present study, we sought to establish and characterize pig retinal primary microglial cell (pMicroglia) culture. We used pig eyes from the local abattoir and optimized pMicroglia cultures using multiple cell culture conditions and methods. The best results were obtained by seeding cells in DMEM-high glucose media for 18 days followed by shaking of the culture plate. The resulting pMicroglia were characterized by cellular morphology, phenotype, and immunostaining with Iba-1, CD68, P2Y12, CD163, CD14, and Isolectin GS-IB4. Generated pMicroglia were found functionally active in phagocytosis assay and responsive to lipopolysaccharides (LPS) in dose-dependent production of IL-1β. Furthermore, they showed increased secretion of pro-inflammatory cytokines with LPS treatment. Thus, we report a novel and reproducible method for the isolation of primary microglial cells from pig eyes, which may be useful for studying retinal diseases.
journal_name
Exp Eye Resjournal_title
Experimental eye researchauthors
Lim RR,Hainsworth DP,Mohan RR,Chaurasia SSdoi
10.1016/j.exer.2019.05.010subject
Has Abstractpub_date
2019-08-01 00:00:00pages
107670eissn
0014-4835issn
1096-0007pii
S0014-4835(18)30546-3journal_volume
185pub_type
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journal_title：Experimental eye research
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journal_title：Experimental eye research
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journal_title：Experimental eye research
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journal_title：Experimental eye research
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journal_title：Experimental eye research
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journal_title：Experimental eye research
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journal_title：Experimental eye research
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更新日期：2015-11-01 00:00:00
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journal_title：Experimental eye research
pub_type： 杂志文章,评审
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更新日期：2014-09-01 00:00:00
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journal_title：Experimental eye research
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更新日期：2007-04-01 00:00:00
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journal_title：Experimental eye research
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journal_title：Experimental eye research
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journal_title：Experimental eye research
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journal_title：Experimental eye research
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journal_title：Experimental eye research
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