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Cited by 433Neuroscience and Neuropharmacology ResearchNeuroinflammation and Neurodegeneration MechanismsNeurogenesis and neuroplasticity mechanisms

Development of a Method for the Purification and Culture of Rodent Astrocytes

Open access
Foo, Lynette C., Allen, Nicola J., Bushong, Eric A., Ventura, P. Britten, Chung, Won-Suk, Zhou, Lu, Cahoy, John D., Daneman, Richard +3 more·2011·Neuron
DOI 10.1016/j.neuron.2011.07.022PMC PMC3172573
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Abstract

The inability to purify and culture astrocytes has long hindered studies of their function. Whereas astrocyte progenitor cells can be cultured from neonatal brain, culture of mature astrocytes from postnatal brain has not been possible. Here, we report a new method to prospectively purify astrocytes by immunopanning. These astrocytes undergo apoptosis in culture, but vascular cells and HBEGF promote their survival in serum-free culture. We found that some developing astrocytes normally undergo apoptosis in vivo and that the vast majority of astrocytes contact blood vessels, suggesting that astrocytes are matched to blood vessels by competing for vascular-derived trophic factors such as HBEGF. Compared to traditional astrocyte cultures, the gene profiles of the cultured purified postnatal astrocytes much more closely resemble those of in vivo astrocytes. Although these astrocytes strongly promote synapse formation and function, they do not secrete glutamate in response to stimulation.

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Crossref · 10.1016/j.neuron.2011.07.022

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  • Endothelial cells secrete heparin-binding EGF-like growth factor, which binds EGFR on astrocytes to promote astrocyte survival.

    “Endothelial cells secrete heparin-binding EGF-like growth factor, which binds its receptor epidermal growth factor receptor on astrocytes to promote astrocyte survival (Foo et al., 2011).”

    From An RNA-Sequencing Transcriptome and Splicing Database of Glia, Neurons, and Vascular Cells of the Cerebral Cortex (2014) · cites this paper as Foo et al., 2011

    In Results, the paper reports HBEGF in endothelial-cell conditioned media and shows that endothelial cells promote astrocyte survival through an EGFR-sensitive effect. It also finds that HBEGF promotes astrocyte survival via EGFR in vitro. But the authors say endothelial-derived HBEGF is *likely* the factor responsible; the claim presents that proposed link as established. (LLM, conf=0.87)

    Experimental — accuracy not yet measured

  • Reducing serum concentration reduces the reactivity of astrocytes and better simulates in vivo conditions.

    “In addition, the serum concentration was gradually reduced and replaced with B-27-supplemented Neurobasal-A medium from DIV4 to DIV6 to reduce the reactivity of astrocytes and better simulate in vivo conditions (30).”

    From Nuclear SphK2/S1P signaling is a key regulator of ApoE production and Aβ uptake in astrocytes (2024) · cites this paper as [30]

    The Results sections on gene expression and calcium imaging report that serum-free, purified astrocyte cultures more closely resemble astrocytes in vivo than traditional cultures. But the paper does not show that gradually reducing serum from DIV4 to DIV6 reduces astrocyte reactivity; its Results section on serum effects says many serum-induced gene-expression changes persisted after serum withdrawal. (LLM, conf=0.84)

    Experimental — accuracy not yet measured

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Cites

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Authors
Foo, L. C., Allen, N. J., Bushong, E. A., Ventura, P. B., Chung, W.-S., Zhou, L., Cahoy, J. D., Daneman, R., Zong, H., Ellisman, M. H., & Barres, B. A.
Affiliations
  • Stanford Medicine (US)
  • University of California San Diego (US)
  • University of Oregon (US)
  • University of California, San Francisco (US)
Corresponding author
Foo, Lynette C.
ORCID
Venue
Neuron
Publisher
Elsevier BV
Published
2011-09
Volume
71
Issue
5
Pages
799-811
ISSN
0896-6273 (ISSN-L)
DOI
10.1016/j.neuron.2011.07.022
PMC
PMC3172573
OpenAlex
W2011331227
Cited by
433
References
5
Citations by year
2026: 19 · 2025: 32 · 2024: 31 · 2023: 44 · 2022: 44 · 2021: 50 · 2020: 33 · 2019: 28 · 2018: 16 · 2017: 25 · 2016: 19 · 2015: 20 · 2014: 22 · 2013: 30 · 2012: 18
FWCI
9.41
Type
journal article
Language
en
Open access
Licence
Locations
  • Neuron (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • National Institutes of Health
  • National Institute of General Medical Sciences (P41GM103412)
  • National Institute of Neurological Disorders and Stroke (R01NS059893)
  • National Center for Research Resources (P41RR004050)
  • Eunice Kennedy Shriver National Institute of Child Health and Human Development (T32HD007348)
Topics
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
  • Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
  • Life Sciences › Neuroscience › Developmental Neuroscience › Neurogenesis and neuroplasticity mechanisms
Keywords
astrocytes, immunopanning, astrocyte culture, postnatal brain, apoptosis, HB-EGF, synaptogenesis, glutamate release, gene expression profiles
MeSH terms
CELF Proteins, Age Factors, Animals, Animals, Newborn, Astrocytes, Cell Count, Cells, Cultured, Cerebral Cortex, Glial Fibrillary Acidic Protein, Membrane Proteins, Neurons, Synapses, RNA-Binding Proteins, Culture Media, Serum-Free, Rats, Sprague-Dawley, Apoptosis, Annexin A5, Receptors, Growth Factor, Gene Expression Regulation, Developmental, Chemokines, Cell Culture Techniques, Intercellular Signaling Peptides and Proteins, Integrin beta Chains, Mice, Rats, Occludin

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