[Effect of different number of bone marrow mesenchymal stem cells on growth of rat dorsal root ganglia in vitro].

OBJECTIVE: Bone marrow mesenchymal stem cells (BMSCs), as replacement cells of Schwann cells, can increase the effect of peripheral nerve repair. However, it has not yet reached any agreement to add the appropriate number of seeded cells in nerve scaffold. To investigate the effect of different number of BMSCs on the growth of rat dorsal root ganglia (DRG).

METHODS: Three 4-week-old Sprague Dawley (SD) rats (weighing 80-100 g) were selected to isolate BMSCs, which were cultured in vitro. Three 1- to 2-day-old SD rats (weighing 4-6 g) were selected to prepare DRG. BMSCs at passage 3 were used to prepare BMSCs-fibrin glue complex. According to different number of BMSCs at passage 3 in fibrin glue, experiment was divided into group A (1 x 10(3)), group B (1 x 10(4)), group C (1 x 10(5)), and group D (0, blank control), and BMSCs were co-cultured with rat DRG. The axon length of DRG, Schwann cell migration distance, and axon area index were quantitatively evaluated by morphology, neurofilament 200, and Schwann cells S-100 immunofluorescence staining after cultured for 48 hours.

RESULTS: Some long cell processes formed in BMSCs at 48 hours; migration of Schwann cells and axons growth from the DRG were observed, growing in every direction. BMSCs in fibrin glue had the biological activity and could effect DRG growth. The axon length of DRG and Schwann cell migration distance in groups A, B, and C were significantly greater than those in group D (P < 0.05). The axon length of DRG and Schwann cell migration distance in group C were significantly less than those in group B (P < 0.05), but there was no significant difference between group A and group C, and between group A and group B (P > 0.05). The axon area index in groups A and B was significantly greater than that in group D (P < 0.05), but there was no significant difference between group C and group D (P > 0.05); there was no significant difference in groups A, B, and C (P > 0.05).

CONCLUSION: In vitro study on DRG culture experiments is an ideal objective neural model of nerve regeneration. The effect of different number of BMSCs in fibrin glue on the growth of DRG has dose-effect relationship. It can provide a theoretical basis for the appropriate choice of the BMSCs number for tissue engineered nerve.