Leukemia inhibitory factor (LIF) is an important factor regulating cell growth, proliferation and differentiation. Bone marrow mesenchymal stem cells (BM-MSCs) have broad application prospects in the repair of tissue damage as well as the treatment of autoimmune diseases, and, as a result, have great potential in the disease treatment and genetic conservation of rare and endangered animals, such as giant pandas. The aim of this study was to construct a giant panda LIF overexpression vector and transfect panda bone marrow mesenchymal stem cells (PDBM-MSCs) with liposome to study the effect of LIF on the growth of PDBM-MSCs. The CDS region of the LIF gene was amplified from the cDNA of PDBM-MSCs, and seamlessly cloned onto the pCDH-CMV-MCS-EF1-GFP+puro overexpression vector to construct the LIF overexpression recombinant plasmid. Lipo3000 was used to transfect recombinant plasmid into PDBM-MSCs. The results showed that PDBM-MSCs expressed green fluorescence 48 h after transfection, and total RNA was extracted and reverse-transcribed into cDNA 72 h after transfection. A q-PCR detection showed that the mRNA expression level of LIF in the LIF overexpression group was significantly higher than that in the negative control group. Moreover, overexpression of LIF could significantly up-regulate the mRNA expressions of CCNB2 and CDK7, and significantly down-regulate the mRNA expressions of apoptosis genes caspase3, P53, P16 and P21. 72 h after transfection, flow detection showed that the viability of PDBM-MSCs transfected with LIF overexpression recombinant plasmid was significantly higher than that of the negative control group. In this study, the panda LIF overexpression vector was successfully constructed and transfected into PDBM-MSCs, which significantly improved cell viability and inhibited apoptosis, indicating that LIF can promote growth and inhibit apoptosis of PDBM-MSCs, laying a foundation for the study of the mechanism of LIF in giant pandas.