Mesenchymal stem cells (MSCs) are multi-potential cells that are able to proliferate and differentiate into other cell types. Much research has been done on the MSCs from the umbilical cord (UCMSCs) in human, mice, and avian, but little literature has been published about these cells in big livestock. Here, we choose Luxi cattle as the experimental animal, we describe an external culture of the UCMSCs from it and summarize the biological characteristics of these cells, e.g., morphologic appearance, surface antigens, colony-forming ability, gene expression, and differentiation potential were detected via using immunofluorescence and reverse transcription polymerase chain reaction (RT-PCR). The induced cells, osteoblast, lipoblast, hepatocyte, islet cells, and neurocyte were identified by Alizarin red staining, Oil-red-O staining, Periodic acid-schiff staining, and Dithizone staining and RT-PCR detection for specific genes. Results suggest that biological characteristics of the UCMSCs were similar to those of MSCs previously analyzed. The primary UCMSCs were sub-cultured to passage 32, the UCMSCs expressed gene CD29, CD44, CD73, CD90, and CD166, induced cells illustrated typical staining, and expressed specific genes, which indicate that the UCMSCs could be a novel alternative source of MSCs for experimental and clinical applications.