Erythropoietin (EPO) can stimulate the proliferation and protraction of endothelial progenitor cells, and plays an important role in the proliferation and differentiation of marrow-derived mesenchymal stem cells (mMSCs) under the acute kidney injury (AKI) microenvironment. In the present study, C57BL/6 mice mMSCs were isolated, and AKI mice models were prepared. The renal cortex was obtained to prepare the ischemia/reperfusion (I/R) kidney homogenate supernatant. P3-mMSCs were treated by different methods: one group was added only I/R kidney homogenate supernatant, and another contained different concentrations of EPO (1, 5, 10, 50 IU/mL) in I/R kidney homogenate supernatant. The proliferation and apoptosis of mMSCs were detected by CCK-8 and TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling), respectively. Expression of erythropoietin receptor (EPOR) and protein of the signal pathway related to proliferation/apoptosis were also examined. The results showed that the proliferation ability of mMSCs treated with I/R kidney homogenate supernatant decreased significantly, while the apoptosis percentage was significantly higher than that of the control. After intervention of EPO, their proliferation enhanced and the apoptosis percentage decreased. EPOR expression was positive in P3-mMSCs. EPO decreased the expression of caspase-3 of mMSCs under the AKI microenvironment in a dose- and time-dependent manner, but increased the Bcl-2 expression. The expression of phosphor-Janus kinase 2, phosphor-signal transducer and activator of transcription (pSTAT-5) increased significantly in 10 IU/mL EPO cultured for five days. Our results show that EPO can promote proliferation of mMSCs in vitro under the AKI microenvironment, which is mediated by EPOR and related with the proliferation/apoptosis signal pathway.