Abstract
Primary cultures of glial and endothelial cells are important tools for basic and translational neuroscience research. Primary cell cultures are usually generated from rodent brain although considerable differences exist between human and rodent glia and endothelial cells. Because many translational research projects aim to identify mechanisms that eventually lead to diagnostic and therapeutic approaches to target human diseases, glia, and endothelial cultures are needed that better reflect the human central nervous system (CNS). Pig brain is easily accessible and, in many aspects, close to the human brain. We established an easy and cost-effective method to isolate and culture different primary glial and endothelial cells from adult pig brain. Oligodendrocyte, microglia, astrocyte, and endothelial primary cell cultures were generated from the same brain tissue and grown for up to 8 weeks. Primary cells showed lineage-specific morphology and expressed specific markers with a purity ranging from 60 to 95%. Cultured oligodendrocytes myelinated neurons and microglia secreted tumor necrosis factor alpha when induced with lipopolysaccharide. Endothelial cells showed typical tube formation when grown on Matrigel. Astrocytes enhanced survival of co-cultured neurons and were killed by Aquaporin-4 antibody positive sera from patients with Neuromyelitis optica. In summary, we established a new method for primary oligodendrocyte, microglia, endothelial and astrocyte cell cultures from pig brain that provide a tool for translational research on human CNS diseases.
Highlights
Glial cells are essential to maintain hemostasis, protect neurons and form myelin in the central nervous system (CNS) (Abbott et al, 2006; Bayraktar et al, 2014; Nave and Werner, 2014; Zou et al, 2014; Domingues et al, 2016; Mosser et al, 2017)
We developed a reliable method to separate microglia, oligodendrocytes, endothelial cells and astrocytes from adult pig brain for primary cultures
Pig brain tissue was dissociated in dissociation buffer to obtain brain cell suspension
Summary
Glial cells are essential to maintain hemostasis, protect neurons and form myelin in the central nervous system (CNS) (Abbott et al, 2006; Bayraktar et al, 2014; Nave and Werner, 2014; Zou et al, 2014; Domingues et al, 2016; Mosser et al, 2017). The expression profile of genes and proteins in human and rodent CNS is quite distinct (Zhang et al, 2016; Galatro et al, 2017) Given these constraints of rodent glia, primary glia cultures from humans or closely related species are considered to be more relevant for investigating human diseases and their treatment. Pig brain tissue is widely and available without the need to kill animals only for research purposes
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