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Peer-reviewedCC BY 4.0STAR Protocols

Protocol for isolation and culture establishment of adult murine neural stem cells using tissue grinder and dissociator-based mechanical dissociation

  • Vanessa Arnold1,2,3,4
  • Anna Wolfram1,2,3,5,6
  • Anastassiya Moskalchuk1,2
  • Vijayasarathy Sampath-Kumar1,2,3,4
  • Abhirami Sunil1,2,3
  • Sogand Ahari1,2,3
  • Lisa Sevenich1,2,4
6 affiliations
  1. 1Department of Neurology and Interdisciplinary Neuro-oncology, Hertie Institute for Clinical Brain Research, University Hospital Tübingen, Tübingen, Germany
  2. 2M3 Research Center for Malignome, Metabolome and Microbiome, University Hospital Tübingen, Tübingen, Germany
  3. 3Graduate Training Centre of Neuroscience, University of Tübingen, Tübingen, Germany
  4. 4Cluster of Excellence iFIT (EXC 2180) ‘Image-Guided and Functionally Instructed Tumor Therapies’, University of Tübingen, Tübingen, Germany
  5. 5Faculty of Pharmacy, University of Castilla-La Mancha, Albacete, Spain
  6. 6Instituto Regional de Investigación Científica Aplicada (IRICA), University of Castilla-La Mancha, Ciudad Real, Spain
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Abstract

Here, we present an enzyme-free workflow for isolating murine adult neural stem cells (aNSCs) using tissue grinder- and dissociator-based mechanical dissociation (TIGR-MD). We describe dissection of the dentate gyrus and subventricular zone, mechanical tissue dissociation, cell recovery, and establishment of primary neurosphere cultures. We further provide procedures for neurosphere maintenance, passaging, and validation. TIGR-MD avoids enzymatic exposure during initial tissue dissociation and supports standardized establishment of primary murine aNSC cultures.

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Cite

BibTeXRIS
BibTeX
@article{arnold2026protocol,
  title = {{Protocol for isolation and culture establishment of adult murine neural stem cells using tissue grinder and dissociator-based mechanical dissociation}},
  author = {Vanessa Arnold and Anna Wolfram and Anastassiya Moskalchuk and Vijayasarathy Sampath-Kumar and Abhirami Sunil and Sogand Ahari and Lisa Sevenich},
  journal = {STAR Protocols},
  year = {2026},
  month = oct,
  doi = {10.1016/j.xpro.2026.104908},
  url = {https://doi.org/10.1016/j.xpro.2026.104908},
  note = {CC BY 4.0},
}
RIS
TY  - JOUR
TI  - Protocol for isolation and culture establishment of adult murine neural stem cells using tissue grinder and dissociator-based mechanical dissociation
AU  - Vanessa Arnold
AU  - Anna Wolfram
AU  - Anastassiya Moskalchuk
AU  - Vijayasarathy Sampath-Kumar
AU  - Abhirami Sunil
AU  - Sogand Ahari
AU  - Lisa Sevenich
PY  - 2026
DA  - 2026/10/20
JO  - STAR Protocols
DO  - 10.1016/j.xpro.2026.104908
UR  - https://doi.org/10.1016/j.xpro.2026.104908
AB  - Here, we present an enzyme-free workflow for isolating murine adult neural stem cells (aNSCs) using tissue grinder- and dissociator-based mechanical dissociation (TIGR-MD). We describe dissection of the dentate gyrus and subventricular zone, mechanical tissue dissociation, cell recovery, and establishment of primary neurosphere cultures. We further provide procedures for neurosphere maintenance, passaging, and validation. TIGR-MD avoids enzymatic exposure during initial tissue dissociation and supports standardized establishment of primary murine aNSC cultures.
SN  - 2666-1667
ER  -