Background: Chondroitin sulfates (CS) is an important glycosaminoglycan’s which is found in sturgeon fishes which are present in cartilage of  these fishes. This compounds, in addition to application in food and cosmetic industries, they have clinical significant  in arthritis treatment and wound healing. Materials and Methods: In present study, the chondroitin sulfate extraction was performed by using pepsin and trypsin digestions with 12 and 18 h interval times followed by cationic salt, cetylprydinium chloride (CPC) treatment. MTT assay at different incubation time (24, 48 and 72 h) was used in order to study of Chondroitin sulfate effect on induction of proliferation of fibroblasts isolated from human skin. In order to determine the proper time and enzymatic digestion, after temperature treatment of samples using pepsin and trypsin enzymes, the SDS-PAGE electrophoresis was used. And the FT-IR analysis was performed to characterize the type of CS. Results: The maximum yield of enzymatic digestion was observed for trypsin digestion at 18h, according to electrophoresis patterns. the amount of CS yield was estimated  at 4.76% in this situation. FT-IR analysis revealed that, the CS belongs to the C4S type. Chondroitin sulfates extracting from cartilage showed concentration-dependent incremental impact on fibroblast cell proliferation so that the dose-response results have shown 167% proliferation at 10 µg/ml dose of CS compared to the control in fibroblast cells. Conclusion: Extracted CS has a positive stimulatory effect on fibroblast cells proliferation in a dose dependent manner. So it can be used for wound healing and some fibroblast related diseases.