Abstract
Human myelopoiesis is an intriguing biological process during which multipotent stem cells limit their differentiation potential generating precursors that evolve into terminally differentiated cells. The differentiation process is correlated with differential gene expression and changes in nuclear architecture. In interphase, chromosomes are distinct entities known as chromosome territories and they show a radial localization that could result in a constrain of inter-homologous distance. This element plays a role in genome stability and gene expression. Here, we provide the first experimental evidence of 3D chromosomal arrangement considering two steps of human normal myelopoiesis. Specifically, multicolor 3D-FISH and 3D image analysis revealed that, in both normal human hematopoietic stem cells and myelod precursors CD14-, chromosomal position is correlated with gene density. However, we observed that inter-homologue distances are totally different during differentiation. This could be associated with differential gene expression that we found comparing the two cell types. Our results disclose an unprecedented framework relevant for deciphering the genomic mechanisms at the base of normal human myelopoiesis.
Highlights
Materials and Methods saline, and CD34+ cells were separated using magnetic cell sorting procedureThe organization of a eukaryotic nucleus reflects its specific expression profile, with dynamic rearrangements and repositioning involved in the regulation of gene expression
cord blood (CB) CD34+ cells were experiments were obtained with a laser optical serial sections was performed using cultured in IMDM added with 20% FCS scanning confocal microscope (LSM 410; voxel-based software algorithms eADS (Bio-Whittaker, Walkersville, MD, USA), in Carl Zeiss MicroImaging) equipped with Ar
The statistical significance s of the inter-homologue and distances was u tested using Student’s t-test. l Analysis of gene expression profiles ia Gene expression profiles of 12 c hematopoietic stem cell (CD34+) and 11 r progenitors (CD14-) samples were obtained e from four different datasets downloaded from gene Expression Omnibus m (Supplementary Table S1)
Summary
Materials and Methods saline, and CD34+ cells were separated using magnetic cell sorting procedureThe organization of a eukaryotic nucleus reflects its specific expression profile, with dynamic rearrangements and repositioning involved in the regulation of gene expression. CB CD34+ cells were experiments were obtained with a laser optical serial sections was performed using cultured in IMDM added with 20% FCS scanning confocal microscope (LSM 410; voxel-based software algorithms eADS (Bio-Whittaker, Walkersville, MD, USA), in Carl Zeiss MicroImaging) equipped with Ar
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