STAR Protocols (Jun 2023)

Mechanical isolation of neonatal and adult mouse dura leukocytes for flow cytometry analysis

  • Chunxiao Niu,
  • Hui Jiang,
  • Mengyao Wu,
  • Tao Zou,
  • Xian Wu,
  • Yuchen Lu,
  • Lijiao Deng,
  • Tingting Guo,
  • Chuan-Yimu Si,
  • Jiyan Zhang

Journal volume & issue
Vol. 4, no. 2
p. 102272

Abstract

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Summary: The meninges, consisting of the pia, arachnoid, and dura layers, provide immunosurveillance of the central nervous system with both innate and adaptive immune cells. Here we present an optimized protocol for isolating dura leukocytes from neonatal and adult mice. We describe steps for harvesting the skull cap, extracting the dura mater, mechanical isolation of dura leukocytes, and flow cytometry analysis. Unlike the time-consuming enzymatic digestion isolation which makes dura hematopoietic stem cells (HSCs) undetectable, this rapid and simplified technique permits dura HSC identification.For complete details on the use and execution of this protocol, please refer to Niu et al. (2022).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

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