Evidence for Diversity in Transcriptional Profiles of Single Hematopoietic Stem Cells 英文参考文献.docVIP

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Evidence for Diversity in Transcriptional Profiles of Single Hematopoietic Stem Cells 英文参考文献.doc

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Evidence for Diversity in Transcriptional Profiles of Single Hematopoietic Stem Cells 英文参考文献

EvidenceforDiversityinTranscriptionalProfiles ofSingleHematopoieticStemCells Carlos A.Ramos1,2,Teresa A.Bowman2,3,Nathan C.Boles2,3,Akil A.Merchant1,2,Yayun Zheng2,Irma Parra4, Suzanne A.W. Fuqua4,Chad A.Shaw5,Margaret A.Goodell2,3,6* 1DepartmentofMedicine,BaylorCollegeofMedicine,Houston,Texas,UnitedStatesofAmerica,2CenterforCellandGeneTherapy,BaylorCollegeofMedicine,Houston, Texas,UnitedStatesofAmerica,3CellandMolecularBiologyProgram,BaylorCollegeofMedicine,Houston,Texas,UnitedStatesofAmerica,4BreastCenter,BaylorCollege of Medicine, Houston, Texas, United States of America, 5 Human and Molecular Genetics, Baylor College of Medicine, Houston, Texas, United States of America, 6 DepartmentofPediatrics,BaylorCollegeofMedicine,Houston,Texas,UnitedStatesofAmerica Hematopoieticstemcellsreplenishallthecellsofthebloodthroughoutthelifetimeofananimal.Althoughthousands ofstemcellsresideinthebonemarrow,onlyafewcontributetobloodproductionatanygiventime.Nothingisknown about the differences between individual stem cells that dictate their particular state of activation readiness. To examinesuchdifferencesbetweenindividualstemcells,wedeterminedtheglobalgeneexpressionprofileof12single stemcellsusingmicroarrays.Weshowedthatatleasthalfofthegeneticexpressionvariabilitybetween12singlecells profiled was due to biological variation in 44% of the genes analyzed. We also identified specific genes with high biologicalvariancethatarecandidatesforinfluencingthestateofreadinessofindividual hematopoieticstemcells, andconfirmedthevariabilityofasubsetofthesegenesusingsingle-cellreal-timePCR.Becauseapparentvariationof somegenesislikelyduetotechnicalfactors,weestimatedthedegreeofbiologicalversustechnicalvariationforeach gene using identical RNA samples containing an RNA amount equivalent to that of single cells. This enabled us to identifyalargecohortofgeneswithlowtechnicalvariabilitywhoseexpressioncanbereliablymeasuredonthearrays atthesingle-celllevel.Thesedatahaveestablishedthatgeneex