Poly(lactic acid) (PLA) and other aliphatic polyesters containing the unit of lactic acid are very popular biodegradable materials. While the degradation products, lactic acids, have been worried to bring with negative influence on biocompatibility, the focused experimental studies are less reported. This study is aimed at an in vitro examination of cytotoxicity of both L-lactic acid and D,L-lactic acid. Mesenchymal stem cells (MSCs) derived from rat bone marrow are employed to test the cytotoxicity of the lactic acids. Considering that the addition of lactic acids not only introduces lactate groups but also alters medium pH and ion strength, these three candidate effects are examined in a decoupled way by setting different comparison groups. The results confirm that the change of medium pH is the predominant factor. It has also been found that D-lactate is more cytotoxic than L-lactate at high concentrations. Yet, either L-or D,L-lactic acids seem acceptable in most of medical applications, because the cytotoxicity is significant only when the concentrations are as high as 20 mmol/L for both of them.