Directing chondrogenic differentiation of human mesenchymal stem cells (MSCs) is currently a challenging problem in tissue engineering of cartilage. Short-peptide motifs are promising new tools to aid in controlling chondrogenesis. The aim of this study was to investigate whether a short bone morphogenetic protein-2 (BMP-2)-derived peptide (BMP peptide) stimulates chondrogenesis of human MSCs in the absence of other growth factors. A high-throughput pellet culture system was used to rapidly collect biochemical data such as glycosaminoglycan (GAG), total collagen, and DNA content, as well as alkaline phosphatase (AP) activity. Cells cultured with ≥100 μg/mL of the peptide produced 74% of the GAG content that cells cultured with BMP-2 produced. Comparable levels of GAG production were promoted by the peptide and BMP-2 over 4 weeks of culture. However, histology revealed that the peptide promoted a more homogenous distribution of GAG than BMP-2. The BMP peptide directed human MSCs to increase collagen production after 3 weeks, but at significantly lower levels compared to BMP-2. Treatment with BMP-2 resulted in a large increase in hypertrophic markers such as AP activity and gene expression of type X collagen, whereas treatment with the peptide resulted in little-to-no increase in these markers. These results suggest that the BMP peptide could be an effective new tool for cartilage tissue engineering.