title: RNA-stabilized whole blood samples but not peripheral blood mononuclear cells can be stored for prolonged time periods prior to transcriptome analysis.
abstract: Microarray-based transcriptome analysis of peripheral blood as surrogate tissue has become an important approach in clinical implementations. However, application of gene expression profiling in routine clinical settings requires careful consideration of the influence of sample handling and RNA isolation methods on gene expression profile outcome. We evaluated the effect of different sample preservation strategies (eg, cryopreservation of peripheral blood mononuclear cells or freezing of PAXgene-stabilized whole blood samples) on gene expression profiles. Expression profiles obtained from cryopreserved peripheral blood mononuclear cells differed substantially from those of their nonfrozen counterpart samples. Furthermore, expression profiles in cryopreserved peripheral blood mononuclear cell samples were found to undergo significant alterations with increasing storage period, whereas long-term freezing of PAXgene RNA stabilized whole blood samples did not significantly affect stability of gene expression profiles. This report describes important technical aspects contributing toward the establishment of robust and reliable guidance for gene expression studies using peripheral blood and provides a promising strategy for reliable implementation in routine handling for diagnostic purposes.
authors.affiliation: Laboratory of Genomics and Immunoregulation, Life and Medical Sciences Institute (LIMES), University of Bonn, Bonn, Germany.
authors.forename: Svenja
authors.initials: S
authors.lastname: Debey-Pascher
authors.forename: Andrea
authors.initials: A
authors.lastname: Hofmann
authors.forename: Fatima
authors.initials: F
authors.lastname: Kreusch
authors.forename: Gerold
authors.initials: G
authors.lastname: Schuler
authors.forename: Beatrice
authors.initials: B
authors.lastname: Schuler-Thurner
authors.forename: Joachim L
authors.initials: JL
authors.lastname: Schultze
authors.forename: Andrea
authors.initials: A
authors.lastname: Staratschek-Jox
mesh_terms: D000328:Adult; D000368:Aged; D001769:Blood; D016000:Cluster Analysis; D015925:Cryopreservation; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D007963:Leukocytes, Mononuclear; D008297:Male; D008545:Melanoma / Q000097:blood / Q000235:genetics; D008875:Middle Aged; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D020871:RNA Stability; D013997:Time Factors; D055815:Young Adult