A simplified method for isolating highly purified neurons, oligodendrocytes, astrocytes, and microglia from the same human fetal brain tissue.
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.
Jana
Malabendu
M
Department of Neurological Sciences, Rush University Medical Center, Chicago, IL 60612, USA.
Jana
Arundhati
A
Pal
Utpal
U
Pahan
Kalipada
K
D000328:Adult; D001253:Astrocytes / Q000502:physiology*; D001921:Brain / Q000166:cytology* / Q000196:embryology*; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004247:DNA / Q000235:genetics; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D020287:In Situ Nick-End Labeling; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D017628:Microglia / Q000502:physiology*; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000378:metabolism; D019636:Neurodegenerative Diseases / Q000473:pathology; D009474:Neurons / Q000502:physiology*; D009836:Oligodendroglia / Q000502:physiology*; D010587:Phagocytosis / Q000502:physiology; D011247:Pregnancy; D020133:Reverse Transcriptase Polymerase Chain Reaction
D009419:Nerve Tissue Proteins; D004247:DNA; D007770:L-Lactate Dehydrogenase