| Objective:To investigate the method of induction bone marrow stromal cells into GABAergic neurons and the therapeutical effect of their transplantation in animal models of epilepsy,Method:The bone marrow stromal cells were isolated and cultured for 4 passages in sterile condition,and identified by immunocytochemistry;The fourth passage bone marrow stromal cells were then cultured in medium DMEM/F12 which contained 2%B27,bFGF 10ng/ml,RA 0.5μmol/L.After 48 hours,part of them were used to detect Nestin antigen;The remained part were changed medium cultured in DMEM/F12 which contained 2%B27,bFGF 10ng/ml,RA 0.5μmol/L and NT-3 20ng/ml,BDNF 20ng/ml,stained GAD65 after 4 days.Epilepsy model: Lithium chloride(3mEq/kg)was administered ip.After 18-20 h injected pilocarpine.Before the sc injection of pilocarpine 1mg/kg methylscopolamine 30 min before the convulsant to reduce the peripheral consequences of pilocarpine administration.The model were assessed by observation of behavior.We judged the models qualification by observed status epilepticus during acute stage or spontaneous epileptic seizure in chronic phase of 2W.The neural stem cells originated from bone marrow stromal cells and neurotrophic factors were transplanted into the hippocampus of epilepsy models.The therapeutical role was evaluated by frozen section immunohistochemistry and ethology.Results:The purity of the fourth passage bone marrow stromal cells identified by immunocytochemistry was 99%.Most part of cells after induction 72 hours was positive for Nestin and after 7 days the expression of GAD67 possessed about 60%.60%of the rats were obviously observed seizure in 30min after injected pilocarpine.The mortality of epilepsy models was high,only 15～20%survived.The neural stem cells originated from BMSCs survived, migrated and incorporated into the host brain tissue and were distributed along special structure, but there was no obvious change in ethology.Conclusion:Neural stem cells originated from BMSCs could be induced into GABAergic neurons and to some extent had a therapeutical effect in epilepsy. |
