Project description:Purpose: to compare the transcriptomic profile of the two most primitive human EPCR+ stem and CD90+EPCR- progenitor populations. Methods: cells were obtained from human cord-blood and flow-sorted using the surface antigens described. 5 EPCR+ and 4 CD90+EPCR- biological replicates were used to extract total RNA and quality was verified (RIN >8). Libraries were prepared using with 20-25 ng/sample of starting RNA. All samples were sequenced two independent times. Conclusions: this study represents the first analysis of transcriptomes from the two most primitive human stem/progenitor populations, with biologic replicates, generated by RNA-seq technology. The optimized data analysis workflows reported here should provide a framework for comparative investigations of expression profiles of other human blood stem/progenitor populations.
Project description:Cannonical hematopoietic stem cell (HSC) population is phenotypically defined as Lin- CD34+ CD38- CD45RA- CD90+ in human adult bone marrow. We recently identified novel HSC subpopulation defined as CD35+ HSCs. We performed microarray analysi to clarify the characteristics of CD35+ HSCs in comparison with other stem/progenitor populations. As compared to CD35- HSCs and progenitor populations, CD35+ HSCs showed lower expression of cell-cycle- or lineage-affiliated genes.suggesting their dormant or immature state.
Project description:Single cell transcriptomic profiling (sc RNA-seq) of the two Human Hematopoietic Stem Cell Populations from cord blood: 18LinnegCD34posCD133pos (hereafter CBC1:CD34pos ) and 18LinnegCD34negCD133pos HSCs (hereafter CBC3:CD34neg). Purpose: to compare the single cells transcriptomic profiles of the two Human Hematopoietic Stem Cell Populations: CD34pos HSCs and CD34neg HSCs in order to find unique homing molecules profile. Results provide insight of highly expressed adhesion molecules in CD34neg HSCs and their crucial role in interacting with the bone marrow microenvironment.
Project description:There are no described quality assurance mechanisms for newly formed stem cells. We observed intimate interactions between macrophages and blood stem cells in zebrafish embryos. Stressed stem cells were marked by surface Calreticulin, which stimulates macrophage interaction as an eat me signal. Macrophage-stem cell interactions either lead to removal of cytoplasmic material and stem cell proliferation or resulted in complete stem cell engulfment. Calreticulin knock down or embryonic macrophage depletion reduced the number of stem cell clones into adulthood. Our work supports a model in which embryonic macrophages determine hematopoietic clonality by monitoring stem cell quality.
Project description:Single cell RNA sequencing of 37,501 cells representing four independently isolated hematopoietic stem and progenitor populations from mouse bone marrow
Project description:Several individual miRNAs (miRs) have been implicated as potent regulators of important processes during normal and malignant hematopoiesis. In addition, many miRs have been shown to fine-tune intricate molecular networks, in concert with other regulatory elements. In order to study hematopoietic networks as a whole, we first created a comprehensive map of global miR expression during early murine hematopoiesis. Next, we determined the copy number per cell for each miR in each of the examined stem and progenitor cell types. As data is emerging indicating that miRs function robustly mainly when they are expressed above a certain threshold (~100 copies per cell), our database provides a resource for determining which miRs are expressed at a potentially functional level in each cell type. Finally, we combine our miR expression map with matched mRNA expression data and external prediction algorithms, using a Bayesian modeling approach to create a global landscape of predicted miR-mRNA interactions within each of these hematopoietic stem and progenitor cell subsets. This approach identifies several interaction networks comprising a “stemness” signature in the most primitive hematopoietic stem cell (HSC) populations, as well as “myeloid” patterns associated with two branches of myeloid development. Six populations of mouse hematopoietic stem and progenitors have been analyzed (LT-HSC, ST-HSC, MPP, CMP, GMP, MEP). 2-3 replicates are included for each sample.