Premature cellular senescence in isolated chondrocytes from polydactyl cartilage in FBS containing serum in comparison to human serum

Abstract

Background. Transplantation of chondrocytes is one of the methods for treatment of articular cartilage defects. For this purpose, it is necessary to isolate these chondrocytes and expand in serum containing culture medium. Sera contain growth factors that are added to medium. Fetal bovine serum (FBS) is used for standard cell culture. However bovine proteins of FBS are accompanied with risk of host immune system reactions. Therefore, it is necessary to replace FBS with human serum for clinical cases. In addition, infants have chondrocytes with high proliferation potency versus adults. In this study, the effects of FBS and human placental serum was investigated on morphology and cellular senescence related genes expression of chondrocytes of infants. Methods. Chondrocytes after isolation of from polydactyly cartilage tissue samples, were cultured in 10% FBS and human serum up to passage 5. Human placental serum was obtained from venous blood of embryonic surface of human placenta. Chondrocytes in two culture medium were compared morphologically by inverted microscope. For cellular senescence evaluation, relative expression of IGF1 and P16 genes were assessed by quantitative real-time RT PCR. Results. Proliferation of chondrocytes is markedly decreased in FBS groups in comparison with human serum. In FBS containing medium, phenotypes of cells were changed from spindle shape to flat appearance after five passages. The expression IGF1 and P16 genes are increased in chondrocytes grown in FBS containing medium and it was significant about P16. Morphologic results were in agreement with Real-time RT PCR results. Conclusion. Human placental serum has advantages such as high proliferation rate on human chondrocytes, prevention from cellular senescence and immunological problems in using cultured chondrocytes for cartilage tissue engineering.