دورية أكاديمية
أعرض في EDS

Gene expression profiling of immunomagnetically separated cells directly from stabilized whole blood for multicenter clinical trials.

التفاصيل البيبلوغرافية
العنوان: Gene expression profiling of immunomagnetically separated cells directly from stabilized whole blood for multicenter clinical trials.
المؤلفون: Letzkus M; Biomarker Development, Novartis Institutes for BioMedical Research (NIBR), Basel, Switzerland., Luesink E; Biomarker Development, Novartis Institutes for BioMedical Research (NIBR), Basel, Switzerland., Starck-Schwertz S; Biomarker Development, Novartis Institutes for BioMedical Research (NIBR), Basel, Switzerland., Bigaud M; Biomarker Development, Novartis Institutes for BioMedical Research (NIBR), Basel, Switzerland., Mirza F; Scientific Capability Development, Pharma-Development, Novartis Pharma AG, Basel, Switzerland., Hartmann N; Biomarker Development, Novartis Institutes for BioMedical Research (NIBR), Basel, Switzerland., Gerstmayer B; Miltenyi Biotec GmbH, Bergisch Gladbach, Germany., Janssen U; Miltenyi Biotec GmbH, Bergisch Gladbach, Germany., Scherer A; Spheromics, Kontiolahti, Finland., Schumacher MM; Biomarker Development, Novartis Institutes for BioMedical Research (NIBR), Basel, Switzerland., Verles A; Biomarker Development, Novartis Institutes for BioMedical Research (NIBR), Basel, Switzerland., Vitaliti A; Biomarker Development, Novartis Institutes for BioMedical Research (NIBR), Basel, Switzerland., Nirmala N; Biomarker Development, Novartis Institutes for BioMedical Research (NIBR), Cambridge, MA, USA., Johnson KJ; Biomarker Development, Novartis Institutes for BioMedical Research (NIBR), Cambridge, MA, USA., Staedtler F; Biomarker Development, Novartis Institutes for BioMedical Research (NIBR), Basel, Switzerland.
المصدر: Clinical and translational medicine [Clin Transl Med] 2014 Nov 13; Vol. 3, pp. 36. Date of Electronic Publication: 2014 Nov 13 (Print Publication: 2014).
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: Wiley Country of Publication: United States NLM ID: 101597971 Publication Model: eCollection Cited Medium: Print ISSN: 2001-1326 (Print) Linking ISSN: 20011326 NLM ISO Abbreviation: Clin Transl Med Subsets: PubMed not MEDLINE
أسماء مطبوعة: Publication: 2020- : [Hoboken, NJ] : Wiley
Original Publication: Heidelberg : Springer-Verlag
مستخلص: Background: Clinically useful biomarkers for patient stratification and monitoring of disease progression and drug response are in big demand in drug development and for addressing potential safety concerns. Many diseases influence the frequency and phenotype of cells found in the peripheral blood and the transcriptome of blood cells. Changes in cell type composition influence whole blood gene expression analysis results and thus the discovery of true transcript level changes remains a challenge. We propose a robust and reproducible procedure, which includes whole transcriptome gene expression profiling of major subsets of immune cell cells directly sorted from whole blood.
Methods: Target cells were enriched using magnetic microbeads and an autoMACS® Pro Separator (Miltenyi Biotec). Flow cytometric analysis for purity was performed before and after magnetic cell sorting. Total RNA was hybridized on HGU133 Plus 2.0 expression microarrays (Affymetrix, USA). CEL files signal intensity values were condensed using RMA and a custom CDF file (EntrezGene-based).
Results: Positive selection by use of MACS® Technology coupled to transcriptomics was assessed for eight different peripheral blood cell types, CD14+ monocytes, CD3+, CD4+, or CD8+ T cells, CD15+ granulocytes, CD19+ B cells, CD56+ NK cells, and CD45+ pan leukocytes. RNA quality from enriched cells was above a RIN of eight. GeneChip analysis confirmed cell type specific transcriptome profiles. Storing whole blood collected in an EDTA Vacutainer® tube at 4°C followed by MACS does not activate sorted cells. Gene expression analysis supports cell enrichment measurements by MACS.
Conclusions: The proposed workflow generates reproducible cell-type specific transcriptome data which can be translated to clinical settings and used to identify clinically relevant gene expression biomarkers from whole blood samples. This procedure enables the integration of transcriptomics of relevant immune cell subsets sorted directly from whole blood in clinical trial protocols.
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فهرسة مساهمة: Keywords: Cell sorting; Clinical; Transcriptomics
تواريخ الأحداث: Date Created: 20150519 Date Completed: 20150518 Latest Revision: 20200928
رمز التحديث: 20221213
مُعرف محوري في PubMed: PMC4424390
DOI: 10.1186/s40169-014-0036-z
PMID: 25984272
قاعدة البيانات: MEDLINE
الوصف
تدمد:2001-1326
DOI:10.1186/s40169-014-0036-z