Document Detail

Isolation of subsets of immune cells.
MedLine Citation:
PMID:  15937347     Owner:  NLM     Status:  MEDLINE    
Subsets of immune cells can be isolated before analysis by the enzyme-linked immunospot (ELISPOT) assay with various cell separation techniques. This chapter describes techniques to select desired cells or deplete unwanted cells by crosslinking cells to dense or magnetic particles for subsequent separation. The RosetteSep method can be used to isolate specific cell types directly from human whole blood, using the red blood cells (RBCs) present in the sample as dense particles. Unwanted cells are crosslinked to multiple RBCs, forming "rosettes." The rosettes, free RBCs, and granulocytes pellet when the sample is centrifuged over a buoyant density medium. The unlabeled, desired cells are simply collected from the interface between the plasma and the buoyant density medium. The SpinSep method for isolation of mouse spleen or bone marrow cells is similar to RosetteSep, except that the unwanted cells are bound to dense particles rather than RBCs. The EasySep immunomagnetic system can be used with cell suspensions from a variety of species. Cells are crosslinked to nanometer-sized paramagnetic particles. Magnetically labeled cells are separated from unlabeled cells by placing the sample in a high gradient magnetic field. Both the labeled and the unlabeled fractions can be recovered for further use.
Carrie E Peters; Steven M Woodside; Allen C Eaves
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  302     ISSN:  1064-3745     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2005  
Date Detail:
Created Date:  2005-06-06     Completed Date:  2005-07-11     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  95-116     Citation Subset:  IM    
StemCell Technologies Inc., Vancouver, British Columbia, Canada.
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MeSH Terms
Antibodies, Bispecific
Cell Separation / methods*
Cross-Linking Reagents
Enzyme-Linked Immunosorbent Assay / methods
Immunomagnetic Separation / methods
Leukocytes / classification,  cytology*,  immunology*
Rosette Formation / methods
Reg. No./Substance:
0/Antibodies, Bispecific; 0/Cross-Linking Reagents

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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