We describe a method to prepare postnatal rat brain primary cell cultures composed of astrocytes, oligodendrocytes, and microglia. After 1 week in vitro, the mixed glial cell cultures are free of neurons, meningeal cells and fibroblasts. We developed a simple procedure to selectively harvest enriched populations of each of the three major glial cell types. Because these cells are at a progenitor/immature stage, each can be further cultured separately in serum or serum-free media to yield large quantities of the desired glial cell subpopulations with a high degree of purity in the range of 96-99%. These cell culture models have been used extensively for performing biochemical, molecular, and pharmacological studies using standard assays and obtain sound quantitative data. These studies have given us insights into the development, properties, and functions of rat and mouse glial cells in vitro. The findings have largely been validated and extended in animal models over the last 3 decades. Since this method has been cited in more than 2,500 research papers, the data obtained across laboratories can be compared more readily.
[Show abstract][Hide abstract] ABSTRACT: Multiple sclerosis (MS) is widely considered to be the result of an aggressive autoreactive T cell attack on myelin. How these autoimmune responses arise in MS is unclear, but they could result from virus infections. Thus, viral and autoimmune diseases in animals have been used to investigate the possible pathogenic mechanisms operating in MS. The autoimmune model, experimental autoimmune encephalomyelitis, is the most widely-used animal model and has greatly influenced therapeutic approaches targeting autoimmune responses. To investigate demyelination and remyelination in the absence of the adaptive immune response, toxin-induced demyelination models are used. These include using cuprizone, ethidium bromide and lysolecithin to induce myelin damage, which rapidly lead to remyelination when the toxins are withdrawn. The virus models include natural and experimental infections such as canine distemper, visna infection of sheep, and infection of non-human primates. The most commonly used viral models in rodents are Semliki Forest virus and Theiler's murine encephalomyelitis virus. The viral and experimental autoimmune encephalomyelitis models have been instrumental in the understanding of how viruses trigger inflammation, demyelination and neurodegeneration in the central nervous system. However, due to complexity of the animal models, pathological mechanisms are also examined in central nervous system cell culture systems including co-cultures, aggregate cultures and brain slice cultures. Here we critically review in vitro and in vivo models used to investigate MS. Since knowledge gained from these models forms the basis for the development of new therapeutic approaches for MS, we address the applicability of the models. Finally, we provide guidance for using and reporting animal studies with the aim of improving translational studies to the clinic.
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