In order to obtain MicroRNA (miRNAs) expression profiles of human hematopoietic stem cells (HSCs), and to preliminarily investigate functions of HSC-correlative miRNAs, by using miniMACS magnetic beads and fluorescence-activated cell sorting (FACS), isolated hematopoietic stem cells (HSCs) were isolated from human umbilical cord blood and performed total RNA extraction. Next, using a microarray, miRNA gene expression profiles of HSCs were obtained. CFC assays were performed to research on differentiation-promoting function of miR-520h, enriched in HSCs. Results showed that CD34⁺ hematopoietic cells and HSCs were successfully isolated from human umbilical cord blood, and 31 HSCs-correlative miRNAs were screened by microarrray. Among them, 22 were low in HSCs, and 9 were high. The result of real time RT-PCR confirmed high expression of miR-520h in HSCs. CFC assays showed that miR-520h promotes differentiation of HSCs into progenitor cells. In conclusion, human HSCs have a set of specific miRNAs that contribute to regulation of HSCs functions, which pave the way for exploring the roles of miRNAs in development of hematopoiectic system.