Despite progress in identifying the cellular composition of hematopoietic stem/progenitor cell (HSPC) niches, little is known about the molecular requirements of HSPC support. To address this issue, we used a panel of recognized HSPC-supportive stromal lines and less-supportive counterparts originating from embryonic and adult hematopoietic sites. Through comprehensive transcriptomic meta-analyses, we identified 481 mRNAs and 17 microRNAs organized in a network implicated in paracrine signaling. Further inclusion of 15 stromal samples demonstrated that this mRNA subset was predictive of the HSPC support. Our gene set contains most of the known HSPC regulators but also a number of novel ones, such as Pax9 and Ccdc80, as validated by functional studies in zebrafish embryos. In sum, our approach has identified the core molecular network required for HSPC support. These cues together with a searchable web resource will inform ongoing efforts to instruct HSPC ex vivo amplification and formation from pluripotent precursors.