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Astrocytes pp 37-47 | Cite as

Isolation and Culture of Astrocytes from Postnatal and Adult Mouse Brains

  • Yingbo HeEmail author
  • Natalie Taylor
  • Anindya Bhattacharya
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1938)

Abstract

Astrocytes play fundamental roles in development and plasticity of the central nervous system. Dysfunction of astrocytes contributes to most of neurological diseases ranging from neurodegenerative diseases to psychological disorders. To better understand the involvement of astrocytes in both physiological and pathological conditions, and further elucidate their underlying mechanisms, it is critical to establish a reliable isolation and culture method for the cells. In this chapter, we describe the isolation and culture protocols of astrocytes from postnatal and adult mouse brains.

Key words

Astrocytes Isolation Cell culture Postnatal Adult 

References

  1. 1.
    Chung WS, Clarke LE, Wang GX, Stafford BK, Sher A, Chakraborty C, Joung J, Foo LC, Thompson A, Chen C, Smith SJ, Barres BA (2013) Astrocytes mediate synapse elimination through MEGF10 and MERTK pathways. Nature 504(7480):394–400CrossRefGoogle Scholar
  2. 2.
    Barres BA (2008) The mystery and magic of glia: a perspective on their roles in health and disease. Neuron 60(3):430–440CrossRefGoogle Scholar
  3. 3.
    Molofsky AV, Krencik R, Ullian EM, Tsai HH, Deneen B, Richardson WD, Barres BA, Rowitch DH (2012) Astrocytes and disease: a neurodevelopmental perspective. Genes Dev 26(9):891–907CrossRefGoogle Scholar
  4. 4.
    Hewett SJ (1999) Interferon-gamma reduces cyclooxygenase-2-mediated prostaglandin E2 production from primary mouse astrocytes independent of nitric oxide formation. J Neuroimmunol 94(1-2):134–143CrossRefGoogle Scholar
  5. 5.
    Uliasz TF, Hamby ME, Jackman NA, Hewett JA, Hewett SJ (2012) Generation of primary astrocyte cultures devoid of contaminating microglia. Methods Mol Biol 814:61–79CrossRefGoogle Scholar
  6. 6.
    de Vellis J, Cole R (2012) Preparation of mixed glial cultures from postnatal rat brain. Methods Mol Biol 814:49–59CrossRefGoogle Scholar
  7. 7.
    McCarthy KD, de Vellis J (1980) Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue. J Cell Biol 85(3):890–902CrossRefGoogle Scholar
  8. 8.
    Marek R, Caruso M, Rostami A, Grinspan JB, Das Sarma J (2008) Magnetic cell sorting: a fast and effective method of concurrent isolation of high purity viable astrocytes and microglia from neonatal mouse brain tissue. J Neurosci Methods 175(1):108–118CrossRefGoogle Scholar
  9. 9.
    Batiuk MY, de Vin F, Duque SI, Li C, Saito T, Saido T, Fiers M, Belgard TG, Holt MG (2017) An immunoaffinity-based method for isolating ultrapure adult astrocytes based on ATP1B2 targeting by the ACSA-2 antibody. J Biol Chem 292(21):8874–8891CrossRefGoogle Scholar
  10. 10.
    Mosher KI, Andres RH, Fukuhara T, Bieri G, Hasegawa-Moriyama M, He Y, Guzman R, Wyss-Coray T (2012) Neural progenitor cells regulate microglia functions and activity. Nat Neurosci 15(11):1485–1487CrossRefGoogle Scholar
  11. 11.
    Zhang Y, Sloan SA, Clarke LE, Caneda C, Plaza CA, Blumenthal PD, Vogel H, Steinberg GK, Edwards MS, Li G, Duncan JA 3rd, Cheshier SH, Shuer LM, Chang EF, Grant GA, Gephart MG, Barres BA (2016) Purification and characterization of progenitor and mature human astrocytes reveals transcriptional and functional differences with mouse. Neuron 89(1):37–53CrossRefGoogle Scholar
  12. 12.
    Chai H, Diaz-Castro B, Shigetomi E, Monte E, Octeau JC, Yu X, Cohn W, Rajendran PS, Vondriska TM, Whitelegge JP, Coppola G, Khakh BS (2017) Neural circuit-specialized astrocytes: transcriptomic, proteomic, morphological, and functional evidence. Neuron 95(3):531–549.e539CrossRefGoogle Scholar
  13. 13.
    Kantzer CG, Boutin C, Herzig ID, Wittwer C, Reiss S, Tiveron MC, Drewes J, Rockel TD, Ohlig S, Ninkovic J, Cremer H, Pennartz S, Jungblut M, Bosio A (2017) Anti-ACSA-2 defines a novel monoclonal antibody for prospective isolation of living neonatal and adult astrocytes. Glia 65(6):990–1004CrossRefGoogle Scholar
  14. 14.
    Zhang Y, Chen K, Sloan SA, Bennett ML, Scholze AR, O'Keeffe S, Phatnani HP, Guarnieri P, Caneda C, Ruderisch N, Deng S, Liddelow SA, Zhang C, Daneman R, Maniatis T, Barres BA, Wu JQ (2014) An RNA-sequencing transcriptome and splicing database of glia, neurons, and vascular cells of the cerebral cortex. J Neurosci 34(36):11929–11947CrossRefGoogle Scholar
  15. 15.
    Foo LC, Allen NJ, Bushong EA, Ventura PB, Chung WS, Zhou L, Cahoy JD, Daneman R, Zong H, Ellisman MH, Barres BA (2011) Development of a method for the purification and culture of rodent astrocytes. Neuron 71(5):799–811CrossRefGoogle Scholar
  16. 16.
    He Y, Yao X, Taylor N, Bai Y, Lovenberg T, Bhattacharya A (2018) RNA sequencing analysis reveals quiescent microglia isolation methods from postnatal mouse brains and limitations of BV2 cells. J Neuroinflammation 15(1):153–165CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Yingbo He
    • 1
  • Natalie Taylor
    • 1
  • Anindya Bhattacharya
    • 1
  1. 1.Neuroscience Drug DiscoveryJanssen Research and Development, LLCSan DiegoUSA

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