Reduced glutathione effect on the proliferation and neuronal differentiation of bone marrow stromal stem cells

Feng Lin-jie; Gan Hong-quan; Yu Xiang-qian; Yang Jian; Bai Wei-fei; Liu Ying-jie

doi:10.3969/j.issn.2095-4344.2016.45.007

Abstract：BACKGROUND:As bone marrow stromal stem cel s can be easily obtained and have multi-directional differentiation potential, it is an important approach to obtain neural stem cel s for treatment of spinal cord injury through induced differentiation of bone marrow stromal stem cel s. <br> OBJECTIVE:To observe the effects of different concentrations of reduced glutathione on proliferation and neuronal differentiation of bone marrow stromal stem cel s. <br> METHODS:Bone marrow stromal cel s were isolated and cultured by limited dilution method. The cloned bone marrow stromal cel s were stimulated with reduced glutathione at different concentrations (0, 5, 10, 20, 40 mmol/L) respectively. After 72 hours of culture, proliferation of bone marrow stromal cel s was examined by MTT assay. The morphology of bone marrow stromal cel s was observed under inverted microscope and the cel s were identified by immunofluorescence staining of microtubule associated protein-2 and glial fibril ary acidic protein. <br> RESULTS AND CONCLUSION:We found that 5 mmol/L reduced glutathione medium could promote the proliferation of bone marrow stromal cel s (P<0.05), but reduced glutathione at 20 and 40 mmol/L inhibited the cel proliferation (P<0.05). The number and size of cel colonies in the 10 mmol/L reduced glutathione group had no obvious difference from the control group. Reduced glutathione at 10 mmol/L was powerful to induce the neuronal differentiation of bone marrow stromal stem cel s, and most cel s expressed microtubule associated protein-2 and glial fibril ary acidic protein. A random selection of 5 fields of view was conducted and percentage of bone marrow stromal cel s differentiating into neuron-like cel s was calculated as (62.0±4.8)%. These results confirmed that low-concentration (5 mmol/L) reduced glutathione can promote the proliferation of bone marrow stromal cel s, while 10 mmol/L reduced glutathione has a strong induction of bone marrow stromal cel s differentiating into neuron-like cel s.