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RNA-stabilized whole blood samples but not peripheral blood mononuclear cells can be stored for prolonged time periods prior to transcriptome analysis.
Debey-Pascher, Svenja; Hofmann, Andrea; Kreusch, Fatima; Schuler, Gerold; Schuler-Thurner, Beatrice; Schultze, Joachim L; Staratschek-Jox, Andrea.
Afiliación
  • Debey-Pascher S; Laboratory of Genomics and Immunoregulation, Life and Medical Sciences Institute (LIMES), University of Bonn, Bonn, Germany.
J Mol Diagn ; 13(4): 452-60, 2011 Jul.
Article en En | MEDLINE | ID: mdl-21704280
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.
Asunto(s)

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Sangre / Leucocitos Mononucleares / Criopreservación / Análisis de Secuencia por Matrices de Oligonucleótidos / Perfilación de la Expresión Génica / Estabilidad del ARN Tipo de estudio: Guideline Límite: Adult / Aged / Female / Humans / Male / Middle aged Idioma: En Revista: J Mol Diagn Asunto de la revista: BIOLOGIA MOLECULAR Año: 2011 Tipo del documento: Article País de afiliación: Alemania

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Sangre / Leucocitos Mononucleares / Criopreservación / Análisis de Secuencia por Matrices de Oligonucleótidos / Perfilación de la Expresión Génica / Estabilidad del ARN Tipo de estudio: Guideline Límite: Adult / Aged / Female / Humans / Male / Middle aged Idioma: En Revista: J Mol Diagn Asunto de la revista: BIOLOGIA MOLECULAR Año: 2011 Tipo del documento: Article País de afiliación: Alemania