Walter & Eliza Hall Institute, Melbourne, VIC, Australia
Haematopoiesis is the process of blood cell formation from stem cells. Recent evidence through single cell RNA-sequencing (scRNA-seq) and lineage tracing reveals a great deal of heterogeneity in stem and progenitor cells, previously thought to be homogeneous. This has wider implications for our understanding of developmental principles. Many techniques are often destructive of the single cell it seeks to measure. For example, one cannot determine gene expression through scRNA-seq, and measure its fate – without a time machine (current science funding precludes purchase). Our earlier work using clone-splitting established that progenitors are largely ‘imprinted’ for their fate. Using this knowledge, we first index sort clones by FACS, thereby registering the phenotype, then pre-expand clones for a few divisions, prior to testing daughters in multiple assays of fate, and RNA-seq. This way, we are able to capture 3 ‘omics per clone: Transcriptomics, phenomics and fate’omics. Performing this for hundreds of clones allows direct linking of each feature, rather than classical methods which link by inference. Data for DC development and haematopoiesis is presented.
Credits: None available.
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