Molecular profiling of hematopoietic stem cells.
Gene expression profiling using microarrays is a powerful method for studying the biology of hematopoietic stem cells (HSCs). Here, we present methods for activating HSCs with the chemotherapeutic drug 5-Fluorouracil, isolating HSCs from whole bone marrow, and performing microarray analysis. We also discuss quality control criteria for identifying good arrays and bioinformatics strategies for analyzing them. Using these methods, we have characterized the gene expression signatures of HSC quiescence and proliferation and have constructed a molecular model of HSC activation and self-renewal.
Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, TX, USA.
Teresa V
TV
Bowman
Akil A
AA
Merchant
Margaret A
MA
Goodell
D000818:Animals; D002138:Calibration; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D005472:Fluorouracil / Q000494:pharmacology; D020869:Gene Expression Profiling / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods* / Q000592:standards; D011786:Quality Control; D012333:RNA, Messenger / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction
D012333:RNA, Messenger; D005472:Fluorouracil