Mesenchymal stem cells from bone marrow of Rhesus monkey (RhBMSCs) were isolated by density gradient centrifugation, purified by adherence separation, and further identified by phenotype and karyotype analysis. Growth characteristics of RhBMSCs were investigated by observation of cell proliferation and detection of apoptosis. Density gradient centrifugation and adherence separation revealed a simple method to obtain fairly pure RhBMSCs. Fusiform was the most common form of cell under observation, and cell karyotype was normal. Phenotyping assay revealed that the RhBMSCs are highly positive (99.2%) for CD29 when compared with the low positive rates (less than 3.2%) for CD34, CD45 and HLA-DR, which indicated a successful isolation of high purity RhBMSCs and a vigorous activity of cells to proliferate. But the proliferation activity of RhBMSCs gradually decreased following the increasing of cell generations. The methods and results of isolation, expansion, identification and growth characteristics of RhBMSCs were discussed in this paper, which may be helpful to understanding the bone marrow mesenchymal stem cells of human, and may serve as the groundwork for orientated differentiation of RhBMSCs and tissue repairment on experimental animal model of rhesus monkey.