ABSTRACT
BACKGROUND:It is unclear whether serial cel passage in vitro influences the differentiation of bone marrow mesenchymal stem cel s into neural stem cel s. OBJECTIVE:To investigate the effect of cel passage on the differentiation of bone marrow mesenchymal stem cel s into neural stem cel s. METHODS:Rat bone marrow mesenchymal stem cel s were isolated and cultured by the whole bone marrow adherence method. Bone marrow mesenchymal stem cel s at passages 3, 6, 9, 12 were incubated in serum-free medium. After culture for 7 and 14 days, cel biological characterization was observed and differenitaiton ability into neural stem cel s was observed by detecting Nestin expression in cel s using flow cytometry. Then, the cel s were further induced to differentiate and cel multipotential differentiation capacity was detected by measurement of nerve enolase and glial acidic protein expression. RESULTS AND CONCLUSION:Under induction, bone marrow mesenchymal stem cel s at different passages were al differentiated into Nestin-positive neural stem cel s. However, there was a significant difference in differentiation proportion of cel s at different passages (P<0.05). Strongest differentiation ability was found in the passage 6 cel s, with the Nestin expression up to (93.7±2.3)%at 7 days of induction and (96.2±1.8)%at 14 days of induction. The proportion of differentiated cel s at passages 6 and 9 was signfi cantly higher than that at passages 3 and 12. Moreover, adherent cel s were positive for nerve enolase and glial acidic protein. Al these findings indicate that the differentiation of bone marrow mesenchymal stem cel s into neural stem cel s is correlated with cel passage. Cel s at lower or higher passages are both detrimental to cel differentiation.
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BACKGROUND: There are numerous inducers used in inducing bone marrow mesenchymal stem cells (BMMSCs) differentiate into neuron-like cells, however, due to poisonous, most chemical inducers can not be used in human.OBJECTIVE: To investigate the effect of ligustrazine on differentiation of rat BMMSCs into neuron-like cells in vitro, and to search for the optimal inductive concentration.METHODS: After SD rats were anesthetized, bone marrow was obtained from the femoral and tibial bones, centrifuged, and the supernatant was discarded. The extracted cells were cultured in L-DMEM supplemented with 15% fetal bovine serum. The expression of CD44 and CD45 of the 5~(th) passage of BMMSCs were identified by immunocytochemical technique. Serum-free L-DMEM medium contains 1.00, 1.25 and 1.50 g/L ligustrazine concentrations were used to induce the 5~(th) passage of BMMSCs in vitro. Morphology changes of BMMSCs were observed under an inverted phase microscope. Expression of nestin, neuron-specific enolase and glial fibrillary acidic protein were identified by immunocytochemical technique, and the expression ratio of neuron-like cells' surface antigens induced by different concentrations of ligustrazine were compared.RESULTS AND CONCLUSION: ①Most primarily cultured BMMSCs adhered to the wall at 3 days after culture, which proliferated faster after passaged, and the 5~(th) passage of cells were mostly purified into BMMSCs, spread radially or vortex-likely. ②The 5~(th) passage of BMMSCs was positive expressed (98.02±0.81)% CD44, but negative for CD45. ③Neuron-like cells with prominence and bifurcation could be seen after induction. The immunocytochemical method showed that nestin and neuron-specific enolase in most induced cells were positive expressed, especially received a highest ration of neuron-specific enolase expressing in the induced group with 1.25 g/L concentration of ligustrazine. It revealed that ligustrazine can induce BMMSCs differentiated into neuron-like cells, and 1.25 g/L is the optimal inductive concentration.
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Objective To investigate the effects of collateral pathways on prognosis of cerebral infarct patients with prior transient ischemic attack (TIA) and its role in neuroprotective mechanism of TIA. Methods A total of 164 cases of the first ever cerebral infarct patients were consecutively allocated into three groups: A , B and C group according to their age.Three groups were divided into two subgroups respectively based on the absence or presence of prior ipsilateral TIA: A1, A2, B1, B2, C1, C2 group. Neurological dysfunctional scores at admission and 1 month after treatment, barthel index, collateral pathways status, and cerebral infarction volume were evaluated respectively. The relationship between development of collateral pathways and prognosis was assessed at the same time. Results The neurological dysfunctional scores and cerebral infarction volume of patients in A1 group and B1 group were significantly lower than those of A2 group and B2 group (P