Single-cell multiplex RT-qPCR analysis of genes related to migration of murine long-term hematopoietic stem cells during development (65.48)

Abstract

Abstract Hematopoietic stem cells (HSC) follow a defined pattern of migration in development. By E14.5, HSC are found in the fetal liver (FL) and colonize the fetal bone marrow (FBM) by E17.5, where they remain during adult life. The mechanism governing HSC migration from the FL to the FBM is not clearly understood. We performed single-cell multiplex RT-qPCR and FACS analysis of 8 genes involved in HSC migration. We show that the expression of the chemokine receptor Cxcr4 varies across microenvironments and developmental times, while N-cadherin and the calcium receptor Casr show only gene expression in FBM at E17.5. Moreover, VE-cadherin maintains high expression only during fetal development, while the integrin subunit α5 increases its variability after E14.5. The integrin subunits α4 and αL as well as the selectin ligand Psgl1 did not show differences in their expression in single HSCs irrespective of the microenvironment. Our data demonstrate that the expression pattern of phenotypically identical, single HSCs fluctuates as a function of developmental and anatomical microenvironment, an observation that could be overlooked using whole population gene expression analysis. This is the first gene expression comparison of migration-related molecules in fetal tissues across developmental times, enhancing the understanding of HSC migration during development. These findings may have potential relevance in improving BM engraftment of clinically transplanted HSC.