Analysis of gene expression in a complex differentiation hierarchy by global amplification of cDNA from single cells

摘要: Abstract Background: Many differentiating tissues contain progenitor cells that differ in their commitment states but cannot be readily distinguished or segregated. Molecular analysis is therefore restricted to mixed populations cell lines which may also heterogeneous, and the critical differences gene expression might determine divergent development are obscured. In this study, we combined global amplification of mRNA transcripts single with identification developmental potential processed on basis fates sibling from clonal starts. Results We analyzed clones four eight hemopoietic precursor had a variety differentiative potentials; generally each formed identical composition secondary culture. Globally amplified cDNA was prepared individual precursors whose identified by tracking fates. Further samples were terminally maturing, homogeneous populations. Together, represented 16 positions hierarchy. Expression patterns sample set determined for 29 genes known involved growth, differentiation function. The cDNAs bipotent erythroid/megakaryocyte neutrophil/macrophage subtractively hybridized, yielding numerous differentially expressed clones. Hybridization such entire consistent differential Conclusion Tracking reliably identifies taken PCR analysis, demonstrates existence distinct stable commitment. Global cells, fates, yields true representation lineage- stage-specific expression, as confirmed hybridization broad panel probes. results provide first mapping these distinguishes between progenitors different states, generate new insights predictions relevant mechanism, introduce powerful tools unravelling genetic lineage divergence.