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Title: | Microarray analysis of human leucocyte subsets: The advantages of positive selection and rapid purification | Authors: | Lyons, P.A Koukoulaki, M Hatton, A Doggett, K Woffendin, H.B Chaudhry, A.N Smith, K.G.C |
Keywords: | CD14 antigen CD16 antigen CD19 antigen CD4 antigen CD8 antigen microsphere RNA article B lymphocyte blood sampling bone marrow cell CD4+ T lymphocyte CD8+ T lymphocyte cell isolation cell lineage cell selection cell specificity controlled study gene expression profiling genetic transcription human human cell leukocyte microarray analysis monocyte neutrophil purification cell separation cytology DNA microarray flow cytometry gene expression profiling isolation and purification lymphocyte subpopulation methodology Cell Separation Flow Cytometry Gene Expression Profiling Humans Lymphocyte Subsets Microspheres Oligonucleotide Array Sequence Analysis RNA |
Issue Date: | 2007 | Citation: | Lyons, P.A, Koukoulaki, M, Hatton, A, Doggett, K, Woffendin, H.B, Chaudhry, A.N, Smith, K.G.C (2007). Microarray analysis of human leucocyte subsets: The advantages of positive selection and rapid purification. BMC Genomics 8 : 64. ScholarBank@NUS Repository. https://doi.org/10.1186/1471-2164-8-64 | Rights: | Attribution 4.0 International | Abstract: | Background: For expression profiling to have a practical impact in the management of immune-related disease it is essential that it can be applied to peripheral blood cells. Early studies have used total peripheral blood mononuclear cells, and as a consequence the majority of the disease-related signatures identified have simply reflected differences in the relative abundance of individual cell types between patients and controls. To identify cell-specific changes in transcription it would be necessary to profile purified leucocyte subsets. Results: We have used sequential rounds of positive selection to isolate CD4 and CD8 T cells, CD19 B cells, CD14 monocytes and CD16 neutrophils for microarray analysis from a single blood sample. We compared gene expression in cells isolated in parallel using either positive or negative selection and demonstrate that there are no significant consistent changes due to positive selection, and that the far inferior results obtained by negative selection are largely due to reduced purity. Finally, we demonstrate that storing cells prior to separation leads to profound changes in expression, predominantly in cells of the myeloid lineage. Conclusion: Leukocyte subsets should be prepared for microarray analysis by rapid positive selection. © 2007 Lyons et al; licensee BioMed Central Ltd. | Source Title: | BMC Genomics | URI: | https://scholarbank.nus.edu.sg/handle/10635/178000 | ISSN: | 14712164 | DOI: | 10.1186/1471-2164-8-64 | Rights: | Attribution 4.0 International |
Appears in Collections: | Staff Publications Elements |
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