Abstract
Evidence is rapidly growing regarding a role of astroglial cells in the pathogenesis of Alzheimer’s disease (AD), and the hippocampus is one of the important brain regions affected in AD. While primary astroglial cultures, both from wild-type mice and from rodent models of AD, have been useful for studying astrocyte-specific alterations, the limited cell number and short primary culture lifetime have limited the use of primary hippocampal astrocytes. To overcome these limitations, we have now established immortalized astroglial cell lines from the hippocampus of 3xTg-AD and wild-type control mice (3Tg-iAstro and WT-iAstro, respectively). Both 3Tg-iAstro and WT-iAstro maintain an astroglial phenotype and markers (glutamine synthetase, aldehyde dehydrogenase 1 family member L1 and aquaporin-4) but display proliferative potential until at least passage 25. Furthermore, these cell lines maintain the potassium inward rectifying (Kir) current and present transcriptional and proteomic profiles compatible with primary astrocytes. Importantly, differences between the 3Tg-iAstro and WT-iAstro cell lines in terms of calcium signaling and in terms of transcriptional changes can be re-conducted to the changes previously reported in primary astroglial cells. To illustrate the versatility of this model we performed shotgun mass spectrometry proteomic analysis and found that proteins related to RNA binding and ribosome are differentially expressed in 3Tg-iAstro vs WT-iAstro. In summary, we present here immortalized hippocampal astrocytes from WT and 3xTg-AD mice that might be a useful model to speed up research on the role of astrocytes in AD.
Highlights
Introduction While inAlzheimer’s disease (AD) astrocytes have been historically associated with reactive gliosis andMost of the information about the role of astrocytes in brain pathology has been collected in in vitro experiments on primary cultures
Primary astroglial cultures were first depleted of microglial cells by magnetic-assisted cell sorting (MACS) using anti-CD11b-conjugated microbeads in order to obtain a population of highly purified astrocytes
We recently reported that transforming growth factorβ[2] (TGFβ2) and TGFβ3 were up-regulated at messenger RNA (mRNA) levels in 3Tg-primary astrocytes as compared to WT astroglial cultures[42]
Summary
Introduction While inAlzheimer’s disease (AD) astrocytes have been historically associated with reactive gliosis andMost of the information about the role of astrocytes in brain pathology has been collected in in vitro experiments on primary cultures. Rocchio et al Cell Death and Disease (2019)10:24 variability, short culture lifetime and limited number of cells if cultures are prepared from specific brain areas, such as the hippocampus. To overcome these limitations, immortalized astroglial lines have been proposed. Few attempts have been reported to immortalize astrocytes from hippocampus and from animal models of AD. In this regard, Morikawa et al.[18] have generated immortalized astrocytes from ApoE2, ApoE3 and ApoE4 knock-in mice
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