Faculty Collaboration Database

Characterization of a functionally active primary microglial cell culture from the pig retina. Exp Eye Res 2019 Aug;185:107670

Date

05/20/2019

Pubmed ID

31103710

DOI

10.1016/j.exer.2019.05.010

Scopus ID

2-s2.0-85067560246 (requires institutional sign-in at Scopus site)   10 Citations

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.

Author List

Lim RR, Hainsworth DP, Mohan RR, Chaurasia SS

Author

Shyam S. Chaurasia PhD Associate Professor in the Ophthalmology and Visual Sciences department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Animals
Antigens, CD
Antigens, Differentiation, Myelomonocytic
Biomarkers
Calcium-Binding Proteins
Cell Culture Techniques
Culture Media
Dose-Response Relationship, Drug
Interleukin-1beta
Lipopolysaccharide Receptors
Lipopolysaccharides
Microglia
Phagocytosis
Primary Cell Culture
Receptors, Cell Surface
Receptors, Purinergic P2Y12
Retina
Sus scrofa