Objective To explore the mechanism by which cancer-associated fibroblasts (CAFs) regulate CD13-high expression neutrophil-like myeloid-derived suppressor cell (CD13^hi-nMDSC) migration in pancreatic ductal adenocarcinoma (PDAC), so as to provide potential molecular targets and experimental evidences for immunotherapy in patients. Methods CAFs were isolated and purified from pancreatic cancer tissues of 5 PDAC patients. The phenotype and purity of CAFs were identified by immunofluorescence and flow cytometry. The expression of related factors in CAF was detected by quantitative polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assay (ELISA). CAF conditioned medium and myeloid-derived suppressor cell (MDSC) migration system were constructed by Transwell to observe the migration of MDSCs and to study the specific mechanisms by which the aforementioned cytokines participate in regulating the migration of MDSCs. Results The isolated primary CAFs expressed activation biomarkers fibroblast activation protein (FAP) and α-smooth muscle actin (α-SMA), while the human foreskin fibroblasts (HFFs) of control cells did not express FAP and α-SMA. qPCR results showed that the mRNA expression levels of interleukin 6 (IL-6), monocyte chemotactic protein 1 (MCP-1), and stromal cell-derived factor 1 (SDF-1) in CAFs were higher than those in HFFs (all P＜0.01). The contents of IL-6, MCP-1, and SDF-1 in the CAF culture supernatant were significantly higher than those in the HFF culture supernatant (all P＜ 0.01), and the secretion content increased with the prolongation of culture time. Compared with HFF conditioned medium and regular medium (RPMI 1640), CAF conditioned medium could recruit more total MDSCs and CD13^hi-nMDSCs (all P＜ 0.01). The addition of SDF-1 recombinant protein alone in the culture system could induce the migration of total MDSCs and CD13^hi-nMDSCs, and the addition of SDF-1 neutralizing antibodies or C-X-C motif chemokine receptor 4 (CXCR4) blocking antibodies could significantly reduce the migration of CD13^hi-nMDSCs induced by CAF conditioned medium (all P＜0.01). Although MCP-1 alone could also induce the migration of total MDSCs and CD13^hi-nMDSCs, the number of CD13^hi-nMDSCs migrating was significantly less than that of the SDF-1 experimental group. The IL-6 recombinant protein did not induce the migration of total MDSCs or CD13^hi-nMDSCs. Conclusion CAFs can mediate the migration of total MDSCs and CD13^hi-nMDSCs in PDAC through SDF-1/CXCR4 pathway.