A simplified method for isolating highly purified neurons, oligodendrocytes, astrocytes, and microglia from the same human fetal brain tissue

Neurochem Res. 2007 Dec;32(12):2015-22. doi: 10.1007/s11064-007-9340-y. Epub 2007 Apr 20.

Abstract

Elucidation of the underlying pathogenic mechanisms leading to apoptosis of neurons and oligodendrocytes and activation of microglia and astrocytes in different neurodegenerative and neuroinflammatory disorders remains a challenge in neuroscience. In order to overcome the challenge and find out therapeutic remedies, it is important to study live and death processes in each and every cell type of the brain. Here we present a protocol of isolating highly purified microglia, astrocytes, oligodendrocytes, and neurons, all four major cell types of the CNS, from the same human fetal brain tissue. As found in vivo, these primary neurons and oligodendroglia underwent apoptosis and cell death in response to neurodegenerative challenges. On the other hand, astroglia, and microglia, cells that do not die in neurodegenerative brains, became activated after inflammatory challenge. The availability of highly purified human brain cells will increase the possibility of developing therapies for different neurodegenerative disorders.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Astrocytes / physiology*
  • Brain / cytology*
  • Brain / embryology*
  • Cell Separation / methods*
  • Cell Survival / drug effects
  • Cells, Cultured
  • DNA / genetics
  • Female
  • Humans
  • Immunohistochemistry
  • In Situ Nick-End Labeling
  • L-Lactate Dehydrogenase / metabolism
  • Microglia / physiology*
  • Nerve Tissue Proteins / analysis
  • Nerve Tissue Proteins / metabolism
  • Neurodegenerative Diseases / pathology
  • Neurons / physiology*
  • Oligodendroglia / physiology*
  • Phagocytosis / physiology
  • Pregnancy
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Nerve Tissue Proteins
  • DNA
  • L-Lactate Dehydrogenase