Getting the right cells to the array: Gene expression microarray analysis of cell mixtures and sorted cells

Peter Szaniszlo, Nan Wang, Mala Sinha, Lisa M. Reece, James W. Van Hook, Bruce A. Luxon, James F. Leary

Research output: Contribution to journalArticle

65 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)191-202
Number of pages12
JournalCytometry Part A
Volume59
Issue number2
StatePublished - Jun 2004

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Keywords

  • Cell fixation
  • Cell sorting
  • Cytomics
  • Flow cytometry
  • Gene expression profile
  • Genomics
  • Immunomagnetic separation
  • Microarray
  • Scatterplot
  • Stem/progenitor cells

ASJC Scopus subject areas

  • Hematology
  • Cell Biology
  • Pathology and Forensic Medicine
  • Biophysics
  • Endocrinology

Cite this

Szaniszlo, P., Wang, N., Sinha, M., Reece, L. M., Van Hook, J. W., Luxon, B. A., & Leary, J. F. (2004). Getting the right cells to the array: Gene expression microarray analysis of cell mixtures and sorted cells. Cytometry Part A, 59(2), 191-202.