Document Detail


Getting the right cells to the array: Gene expression microarray analysis of cell mixtures and sorted cells.
MedLine Citation:
PMID:  15170598     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: Most biological samples are cell mixtures. Some basic questions are still unanswered about analyzing these heterogeneous samples using gene expression microarray technology (MAT). How meaningful is a cell mixture's overall gene expression profile (GEP)? Is it necessary to purify the cells of interest before microarray analysis, and how much purity is needed? How much does the purification itself distort the GEP, and how well can the GEP of a small cell subset be recovered? METHODS: Model cell mixtures with different cell ratios were analyzed by both spotted and Affymetrix MAT. GEP distortion during cell purification and GEPs of purified cells were studied. CD34+ cord blood cells were purified and analyzed by MAT. RESULTS: GEPs for mixed cell populations were found to mirror the cell ratios in the mixture. Over 75% pure samples were indistinguishable from pure cells by their overall GEP. Cell purification preserved the GEP. The GEPs of small cell subsets could be accurately recovered by cell sorting both from model cell mixtures and from cord blood. CONCLUSIONS: Purification of small cell subsets from a mixture prior to MAT is necessary for meaningful results. Even completely hidden GEPs of small cell subpopulations can be recovered by cell sorting.
Authors:
Peter Szaniszlo; Nan Wang; Mala Sinha; Lisa M Reece; James W Van Hook; Bruce A Luxon; James F Leary
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Cytometry. Part A : the journal of the International Society for Analytical Cytology     Volume:  59     ISSN:  1552-4922     ISO Abbreviation:  Cytometry A     Publication Date:  2004 Jun 
Date Detail:
Created Date:  2004-06-01     Completed Date:  2004-12-13     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  101235694     Medline TA:  Cytometry A     Country:  United States    
Other Details:
Languages:  eng     Pagination:  191-202     Citation Subset:  IM    
Copyright Information:
Copyright 2004 Wiley-Liss, Inc.
Affiliation:
Department of Microbiology and Immunology, University of Texas Medical Branch at Galveston, 77555, USA.
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MeSH Terms
Descriptor/Qualifier:
Blood Cells
Cell Line
Cell Separation / methods*
Cells
Flow Cytometry / methods
Gene Expression Profiling*
Humans
Image Processing, Computer-Assisted
Oligonucleotide Array Sequence Analysis / methods*
Grant Support
ID/Acronym/Agency:
5R44RR015374/RR/NCRR NIH HHS; R01-GM38645/GM/NIGMS NIH HHS; R01EB-00245/EB/NIBIB NIH HHS; T32 AI 075261/AI/NIAID NIH HHS

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