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Centrifugal Elutriation for Studies of Neuroimmunity

  • Santhi GorantlaEmail author
  • Myhanh Che
  • Howard E. Gendelman
Protocol
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

Centrifugal elutriation is a technique used to fractionate synchronized populations of cells based on size and density. Notably, countercurrent centrifugal elutriation (CCE) can be used for large cell number isolations while minimally interfering with the cells’ functional properties. This makes it superior to other cell separation techniques such as adherence and fluorescence cell sorting. Separating cells by large scale CCE isolation for monocytes and lymphocytes from whole blood while technically challenging is critical to recapitulate immune responses operative during disease. This includes central nervous system infections and degenerative and inflammatory disorders. Monocytes differentiate rapidly into macrophage-like cells, which can reflect microglial function. Such cells are the primary innate immune effector cells, the sources of immunoregulatory factors, and those that effect clearance of debris. Perhaps, most importantly, they serve as antigen presenters and the target cells for a range of microbial pathogens that include the human immunodeficiency virus while reacting to misfolded, aggregated, and oxidized proteins such as amyloid beta, alpha synuclein, and superoxide dismutase present in Alzheimer and Parkinson’s diseases and amyotrophic lateral sclerosis. They can also readily be examined for their genomic and proteomic structures and functional capacities. Also, it is noteworthy to state that using such unmanipulated cells, reproducible outcomes in an experimental setting reflective of human disease can be performed.

Keywords

Monocytes Monocyte-derived macrophages T cells Leukopak Countercurrent centrifugal elutriation 

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Santhi Gorantla
    • 1
    Email author
  • Myhanh Che
    • 1
  • Howard E. Gendelman
    • 1
  1. 1.Department of Pharmacology and Experimental NeuroscienceUniversity of Nebraska Medical CenterOmahaUSA

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