The Possibility of Microglial and Astroglial Cell Lines Use for Investigation of Preparations for the Therapy of Brain Pathologies
About the author:
Makarenko A. N., Dovgiy R. S.
Heading:
LITERATURE REVIEWS
Type of article:
Scentific article
Annotation:
Microglial cell lines. Over the past few decades, there has been an increased interest in microglia, as many investigators have recognized the importance of this cell in the homeostasis, as well as various pathologies, of the central nervous system. in vitro cultures present a beneficial tool to study the activation state, releasable factors, motility, and other crucial components that characterize microglia, which cannot be sufficiently examined in vivo. Primary microglia cultures. Primary microglia cultures are very prevalent in research due to the similarities in phenotype to in vivo cells. These cells are most often derived from the cortex of a rat or mouse before or early after birth. The advantages of using amoeboid microglia directly from an animal are the functional characteristics that these cells are endowed with, such as secretory products and cell surface markers, which closely resemble endogenous cells. However, the extensive preparations needed for each experiment makes this model more time consuming and perhaps less attractive as compared to other microglia lines that have shorter prep-time, but maintain similar cell properties. Retroviral immortalized microglia: BV2 and N9. Such immortalized cell lines can be generated by infecting the cells with a retrovirus. Two commonly used cell lines of this type are the BV2 and N9 microglia cell lines which are derived from rat and mouse, respectively. Even with the similarity to primary microglia, the cells do contain oncogenes that render them in some ways different from primary microglia, such as increased proliferation and adhesion, and increased variance of morphologies. The N9 microglia cells were developed by immortalizing murine primary microglia cells with the v-myc or v-mil oncogenes of the avian retrovirus MH2. These cells are genetically modified, which leads to increased proliferation and adherence. Human immortalized microglia: HMO6. it is more difficult to obtain these cells because they have to be derived from human embryos, which can be difficult to access due to the ethical and legal issues. HMO6 cell line was established by using primary human embryonic microglia cultures. HMO6 cells were positive for staining with Ricinus communis agglutinin and CD11b and express transcripts for purinergic receptors, confirming a microglia phenotype. These human-derived cell lines are rarely used based on the fact that it is difficult to obtain a primary culture to transfect. More research is conducted in non-human animals and, thus, murine microglia cultures are more commonly used. Astroglial cell lines. Astrocytes provide a number of supportive functions, such as metabolic support of neurons. A lot of information about astrocytes received from in vitro studies, that’s why cell lines represent a useful tool for investigation of different properties of this type of cells. Similar to microglia, there are primary and immortalized astrocyte cell lines. Primary astrocyte cultures. Most protocols for preparing astroglial-enriched cultures from rat/mouse late embryos/neonates are derived from the seminal work of Booher and Sensenbrenner or the later modification by McCarthy and de Vellis. In none of primary cell cultures do astrocytes represent 100 % of cells. Depending on culture conditions, oligodendrocytes, neurons, various types of precursors, ependymal cells, fibroblasts, endothelial cells or microglial cells can be present in these cultures, generally in small proportions. Immortalized astrocytes. Neu7 cells obtained by infection of rat primary cortical astrocytes with retrovirus containing neu oncogene in its genome. This cell line was made as a model of reactive astrocytes for the screening of factors, which may regulate the growth of neurites under the lesions of central nervous system. Although it is considered, that primary astrocytes promote axonal growth, Neu7 cells inhibit the growth of neurites. This inhibitory effect partially mediated by increased expression of proteoglycan NG2. A7 cell line obtained from rat optical nerve cells, immortalized by transfection of of SV40 large T antigen. Unlike Neu7, these cells promote the growth of neurites. Thus, for investigation of preparations, designed for the therapy of brain pathologies, the most appropriate are the primary glial cell lines, because they are the most similar to the cells in vivo. But, of course, such issues, as presence of different types of cells in astroglial cultures, or time consuming preparation of primary cultures, make them less attractive, than immortalized cell lines.
Tags:
glia, microglia, astrocytes, cell line.
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Publication of the article:
«Bulletin of problems biology and medicine» Issue 4 part 1 (113), 2014 year, 19-24 pages, index UDK 57. 085. 23