Isolation, characterization, and cultivation of astrocytes

  • Isolation of pure and viable astrocytes from neonatal and adult rodent brain in half a day
  • Astrocyte isolation and characterization with novel astrocyte-specific antibodies
  • Cultivation of primary astrocytes in serum-free medium

MACS Academy: Astrocyte research – novel tools for characterization, isolation, and cultivation

Melanie Jungblut gives an overview of novel approaches for the characterization and isolation of astrocytes and neural stem cells by combination of optimized cell preparation methods and sophisticated cell sorting technologies. Topics include unique astrocyte-specific antibodies, a novel approach for gentle sorting of adult neural stem cells with the MACSQuant® Tyto® Cell Sorter, how to optimize neonatal and adult brain tissue dissociation, and improved serum-free cultivation of primary astrocytes and adult neural stem cells.

Application data by workflow step

Murine brain tissue dissociation

Purity of cell suspensions before and after sample clearing
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Pure cell suspensions from adult brain

The Adult Brain Dissociation Kit helps to remove  cell debris and erythrocytes to acquire clean single cell suspensions for downstream isolation of astrocytes.

Purity and viability of cells obtained from murine (P4) whole brain tissue
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Highly viable neonatal astrocyte cell suspensions

The gentle dissociation of P4 mouse brain tissue with the Neural Tissue Dissociation Kit (P) results in a high percentage of viable cells.

Magnetic isolation of astrocytes

Flow cytometry analysis of dissociated P3 mouse brain before and after neonatal astrocyte isolation
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Pure neonatal astrocytes in one hour

The MACS® MicroBead Technology in combination with Neural Tissue Dissociation Kits allows the isolation of pure neonatal astrocytes in one hour. Using astrocyte-specific MicroBeads based on the novel anti-GLAS (ACSA-1) or anti-ACSA-2 monoclonal antibodies results in purities over 90%.

Flow cytometry analysis of dissociated adult brain before and after adult astrocyte isolation
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Pure adult astrocytes in one hour

Highly pure and viable adult astrocytes can be isolated from single-cell suspensions obtained with the Adult Brain Dissociation Kit, mouse and rat by using the Anti-ACSA-2 MicroBead Kit, mouse.

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How to isolate pure viable astrocytes

Want to know the tips and tricks of how to get pure and viable astrocytes from adult brain in half a day? In our new video tutorial, our astrocytes expert from R&D shows you the entire workflow step by step, including:

  • Automated dissociation of adult rodent brain tissue using the Adult Brain Dissociation Kit and gentleMACS™ Octo Dissociator with Heaters
  • Isolation of pure, viable, and functional adult astrocytes from mouse brain in half a day, using MACS® Technology
  • Downstream characterization and culture of isolated cells
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Scientific poster
Efficient isolation of viable primary neural cells from adult murine brain tissue based on a novel automated tissue dissociation protocol

Hui Zhang1, Sandy Reiß1, Stefan Tomiuk1, Silvia Rüberg1, Richard Fekete2, Melanie Jungblut1, and Andreas Bosio1

1Miltenyi Biotec B.V. & Co. KG, Bergisch Gladbach, Germany; 2Fluidigm Corporation, South San Francisco, CA, USA

Astrocyte cultivation in serum-free astrocyte medium

Astrocyte growth with different seeding densities
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Superior astrocyte growth even with low seeding densities

Astrocytes cultured in AstroMACS Medium show an increased expansion rate. Hence, superior astrocyte growth is achieved even when seeded with low density.

Neonatal astrocytes cultured in AstroMACS Medium
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Preservation of healthy astrocytes for downstream experiments

Neonatal astrocytes cultured in AstroMACS Medium show healthy astrocyte morphology
and cell-cell contacts.

Adult astrocytes cultured with or without using AstroMACS Separation Buffer
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Less dead cells and enhanced adult astrocyte culture

AstroMACS Medium allows enhanced adult astrocyte culture together with the new AstroMACS Separation Buffer, which removes dead cells during cell separation and provides a healthy culture environment.

Related PDFs:

AstroMACS Medium (brochure) 

Astrocyte-specific immunostaining and microscopy analysis

Co-expression of ACSA-2 and GLAST in cultured astrocytes and mouse brain sections
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Astrocyte-specific antibodies

Specific antibodies are essential for reliable neural cell studies, especially for cell types with high diversity, such as astrocytes. The monoclonal ACSA-2 and GLAST antibodies are pan antibodies and recognize both, non-reactive and reactive astrocytes.


  • Jungblut,  M.  et  al.  (2012)  Isolation  and  characterization  of  living  primary astroglial  cells  using  the  new  GLAST-specific  monoclonal  antibody  ACSA-1. Glia 60 (6): 894–907.
  • G. Kantzer, C. et al. (2017) Anti-ACSA-2 defines a novel monoclonal antibody for prospective isolation of living neonatal and adult astrocytes. Glia 65(6): 990–1004.
  • Batiuk, M. Y. et al. (2017) An immunoaffinity-based method for isolating ultrapure adult astrocytes based on ATP1B2 targeting by the ACSA-2 antibody. J. Biol. Chem. 292(21): 8874–8891.

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