Objective: To study the possibility of primary human chondrocytes culture in gelatin scaffold and the effects of exogenous HA on chondrocyte differentiation and synthesis of the hyaline-like extracellular matrix. Material and Method: Cartilage tissue was engineered by using primary human chondrocytes with HA-treated gelatin scaffolds and gelatin scaffolds. The chondrogenic properties were monitored for chondrocyte proliferation, adhesion, and hyaline-like extracellular matrix production in both groups. The results were compared to each other. Results: Chondrocyte proliferation, adhesive activity, and new HA production were significantly increased in HA-treated gelatin scaffold (p < 0.05). Immuno histochemistry for WF6 epitope demonstrated the higher quality of hyaline-like extracellular matrix production. Moreover, the scanning electron micrograph showed a higher filling of extracellular matrix in the pore of scaffold of HA-treated gelatin scaffold than that in non-HA treated scaffold. Conclusion: The present study demonstrated the possible role of commercial gelatin-based scaffold in cartilage tissue engineering. It also demonstrated that exogenous HA-treated scaffold provides positive effects for chondrocytes.