KHẢO SÁT KHẢ NĂNG TĂNG SINH, DI CƯ VÀ BIỆT HOÁ CỦA TẾ BÀO GỐC TRUNG MÔ TỦY XƯƠNG NGƯỜI SAU BẢO QUẢN ĐÔNG LẠNH

Abstract

Objective: To evaluate the proliferation, migration and differentiation capacity of bone marrow stem cells (BMSCs) after cryopreservation. Method: Alveolar bone from three healthy human subjects was utilised to isolate and culture BMSCs. The culturing protocol was employed from the Tissue Engineering and Biomedical Materials laboratory, University of Science, Vietnam National University, Ho Chi Minh City. Subsequently, BMSCs at P3 were cryopreserved in liquid nitrogen for 12 months. Following this period, the BMSCs were thawed and cultured to P4 for examination of their proliferation, migration, and differentiation capacity. For the proliferation experiment, BMSC numbers were quantified by measuring optical density at 570nm on days 1, 3, 5, 7, and 9. For the migration experiment, the acellular areas were measured and calculated at 8 hours and 24 hours. For the differentiation experiment, BMSCs were exposed to osteogenic medium in 21 days. Real-time PCR was performed to detect gene expression of RUNX2, COL1A1, and OC. Results: The proliferation experiment results demonstrated that the optical density measured of BMSCs cultured in DMEM/F12 + 10% FBS medium exhibited a gradual increase from day 1, reached its peak at day 5, and subsequently decreased to day 9 (p < 0.001). In the DMEM/F12 group, optical density decreased from day 1 to day 9 (p < 0.001). The migration experiment revealed that the percentage of acellular area reduced at 8 hrs and 24 hrs (p = 0.001). In the differentiation experiment, the expression of RUNX2 and COL1A1 was observed after BMSCs were exposed to osteogenic medium in 21 days. However, no expression of OC was detected. Conclusion: Following cryopreservation, the BMSCs successfully maintain their proliferation, migration and differentiation capacities.