Currently, the methods utilized to describe cell states include defining populations via flow cytometry, examining developmental potentials using in vitro colony-forming-unit assays and in vivo genetic marking, and genomics analyses such as single-cell RNA-Seq (scRNA-Seq) and scATAC-Seq. While these complementary analyses define a multitude of cell states, there is a lack of coherence between the resulting observations partly due to lack of uniform approaches between labs. The end result has been considerable confusion or controversy rather than a consolidation of understanding. To address this fundamental problem in the field we have assembled an interdisciplinary research team which encompasses expertise in the application of single- cell technologies, hematopoiesis, computational genomics and systems biology to develop and promote a unifying framework for the analysis of genomic states. Specifically, based principally on their genomic states, we will define prevalent and rare hematopoietic intermediates, and the optimal markers and flow gates necessary to isolate them. Deliverables will include a consolidated understanding of the hematopoietic hierarchy based on cutting edge technologies.
The proposed work focuses on identifying the transcriptionally coherent stem and progenitor cells encompassing human blood cell production, as well as optimal flow cytometry strategies to isolate them.
