GSE172128
Spatial transcriptome profiling by MERFISH reveals fetal liver hematopoietic stem cell niche architecture [bulk RNA-seq]
The hematopoietic stem cell (HSC) niche has been extensively studied in bone marrow, yet a more systematic investigation into the microenvironment regulation of hematopoiesis in fetal liver is necessary. Here we investigate the spatial organization and transcriptional profile of individual cells in both wild type and Tet2-/- fetal livers, by multiplexed error robust fluorescence in situ hybridization (MERFISH). We find that specific pairs of fetal liver cell types are preferentially positioned next to each other. Ligand-receptor singling molecule pairs such as Kitl and Kit are enriched in neighboring cell types. The majority of HSCs are directly in contact with endothelial cells (ECs) in both wild type and Tet2-/- fetal livers. Loss of Tet2 increases the number of HSCs, and upregulates Wnt and Notch signaling genes in the HSC niche. Two subtypes of ECs – arterial ECs and sinusoidal ECs – and other cell types contribute distinct signaling molecules to the HSC niche. Collectively, this study provides a comprehensive picture and bioinformatic foundation for HSC spatial regulation in fetal liver. Overall design: E14.5 pregnant female C57BL/6 WT mice were used for fetal liver preparation. Total RNA was extracted from E14.5 fetal liver using RNeasy Mini Kit. mRNA was isolated using the Dynabeads mRNA Purification Kit. The cDNA libraries were produced with a NEBNext® Ultra™ II RNA Library Prep Kit for Illumina® and NEBNext® Multiplex Oligos for Illumina® according to the manufacturer's description. Paired-end reads were aligned to the mouse genome (Gencode vM14) with Tophat and transcripts were assembled with Cufflinks.