While several isoforms of parathyroid hormone-related peptide (PTHrP) have been commercially available, the difference in their effect has not been widely studied. The purpose of this study was to determine which isoform most effectively promoted chondrogenesis and suppressed hypertrophy from mesenchymal stem cells (MSCs). MSCs isolated from fresh bone marrow were cultured in pellet in chondrogenic medium containing 5 ng/ml of transforming growth factor (TGF)-β(3). From day 14 of culture, subsets of pellets were additionally treated with one of the four PTHrP isoforms (1-34, 1-86, 7-34, and 107-139) at 100 nM. After a further 2 weeks of in vitro culture, pellets were harvested for analysis. PTHrPs 1-34 and 1-86 significantly decreased the DNA level (p<0.05) while PTHrPs 7-34 and 107-139 significantly increased DNA level (p<0.05) compared with the control treated with TGF-β(3) only. Glycosaminoglycan per DNA significantly increased when treated with PTHrPs 1-34 and 1-86 (p<0.05) while it significantly decreased with PTHrPs 7-34 and 107-139 (p<0.05). PTHrP 1-34 significantly increased the gene and protein expression of the chondrogenic marker COL2A1, and decreased those of hypertrophic markers COL10A1 and alkaline phosphatase while other isoforms showed inconsistent effects. All of PTHrP isoforms significantly suppressed the gene and protein expression of indian hedgehog (p<0.05) while all isoforms except PTHrP 107-139 significantly reduced the gene and protein expression of patched 1 (p<0.05). In conclusion, of several PTHrP isoforms, PTHrP 1-34 most significantly enhanced chondrogenesis and suppressed hypertrophy in MSCs, supporting its use for cartilage tissue engineering.