Culture and identification of human bone marrow mesenchymal stem cells from alveolar ridge dental implant site

摘要

OBJECTIVE: The objective of this study was to establish the culture method of human bone marrow mesenchymal stem cells (BMSCs) from alveolar bone marrow. METHODS: Alveolar bone marrow complex samples were obtained from 35 patients, 22 to 65 years of age, during the course of dental implant treatment by low-speed method. Bone marrow mesenchymal stem cells were seeded and maintained in culture with 10% fetal bovine serum. The form of the cultured cells was observed under inverted microscope, and the cell proliferation capacity was detected. Cell cycle and the antigen expression of P3 BMSCs were measured with flow cytometry. RESULTS: From a small volume (about 0.1-0.2 mL) of bone marrow complex, alveolar BMSCs expanded at a success ratio of 29 (83%) of 35. There is dysmorphism in cultured cells, which mainly were long spindle, polygon, and triangle. Flow cytometry instrument test showed that the cells in G0/G1 phase were an average of 79.29% ± 1.70% and in S phase were an average of 11.09% ± 0.87%. For antibody expression rate: CD29 is 88.00% ± 1.56%, CD44 is 88.15% ± 1.64%, CD34 is 0.42% ± 0.10%, CD45 is 0.45% ± 0.12%, and CD11b is 0.45% ± 0.14%. CONCLUSION: The cells of the marrow complex obtained by low-speed method in implant site preparation cultured in vitro were identified as BMSCs through the morphological observation and the flow cytometry. It is a kind of feasible and simple culture method of human primary BMSCs.