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2022, Vol. 26 ›› Issue (1): 45-51Chondrogenic differentiation of peripheral blood-derived mesenchymal stem cells induced by transforming growth factor beta 3: a dose-effect relationshipYang Tengyun1, Li Yanlin1, Liu Dejian1, Wang Guoliang1, Zheng Zhujun2
Abstract: BACKGROUND: With the development of cartilage tissue engineering technology, there have been in vitro culture and animal experiments on repairing cartilage defects with peripheral blood-derived mesenchymal stem cells, and good progress has been made. Transforming growth factor beta 3 is often used in cell culture in vitro to induce chondrogenic differentiation of mesenchymal stem cells. However, in the experimental application, the dose-effect study of transforming growth factor beta 3 is rarely involved, and the cell culture experiments involving dose-effect often misuse one-way analysis of variance or t-test to analyze this kind of data. OBJECTIVE: To analyze the dose-effect relationship of chondrogenic differentiation potency of peripheral blood-derived mesenchymal stem cells induced by transforming growth factor beta 3 by repeated measurement, in order to obtain a better dosage of transforming growth factor beta 3, and obtain a better effect of chondrogenic differentiation of peripheral blood-derived mesenchymal stem cells. METHODS: After mobilization, extraction and culture of peripheral blood-derived mesenchymal stem cells of Diannan small eared pigs, transforming growth factor beta 3 at different concentrations (25-500 μg/L) was added for chondrogenic differentiation or not added (control group). Cell proliferation in each group was detected by CCK8 assay at 2, 4, 6, 8, 10, 12 and 14 days. At 3, 7, 14 and 21 days of culture, the supernatant of each group was collected, and the level of type II collagen in the supernatant was detected by ELISA. At 21 day of culture, toluidine blue staining was used to observe the expression of the aggregated proteoglycan. At 21 days, immunocytochemical staining was performed to observe the expression of type II collagen. RESULTS AND CONCLUSION: In the range of 25-500 μg/L, the proliferation ability of peripheral blood-derived mesenchymal stem cells was the strongest at the concentration of transforming growth factor beta 3 at 40 μg/L. The concentration of transforming growth factor beta 3 was more likely to promote the chondrogenic differentiation of peripheral blood-derived mesenchymal stem cells in 40 and 80 μg/L. Key words: stem cells, peripheral blood derived mesenchymal stem cells, cytokines, transforming growth factor beta 3, repeated measurements, cartilage tissue engineering, collagen type II, aggrecan, cartilage repair |