骨髓基质干细胞的诱导分化及其治疗癫痫的实验研究  

INDUCED DIFFERENTIATION OF BONE MARROW STROMAL CELLS INTO NEURON-LIKE CELLS AND THEIR TRANSPLANTATION IN ANIMAL MODEL OF EPILEPSY

作　　者：王晓霞[1] 王春芳[1] 李鹏飞[1] 李静[1] 索耀君[1] 

机构地区：[1]山西医科大学医学实验中心,山西太原030001

出　　处：《中国组织化学与细胞化学杂志》2007年第6期637-642,共6页Chinese Journal of Histochemistry and Cytochemistry

基　　金：山西省自然科学基金(20051095)

摘　　要：目的探讨骨髓基质干细胞诱导分化为神经元样细胞的方法及脑内移植治疗大鼠癫痫模型的作用。方法无菌条件下分离纯化BMSCs,用bFGF 10ng/ml、RA0.5μmol/L的DMEM/F12诱导72h后,部分用于免疫荧光检测nestin,其余的继续用bFGF 10ng/ml、RA 0.5μmol/L及神经营养因子NT-3 20ng/ml、BDNF 20ng/ml的DMEM/F12诱导4d,检测GAD67。皮下注射匹罗卡品建立癫痫大鼠模型,采用行为学分析筛选模型。通过立体定位仪,用微量注射器将BMSCs来源的神经干细胞和神经营养因子移植入癫痫鼠海马内,观察大鼠行为变化,存活2、4周后,心脏灌注取脑,冰冻切片免疫组化双标检测移植细胞的存活、迁移、分化情况。结果BMSCs诱导72h后,nestin表达阳性,4d后GAD67检测60%阳性。采用匹罗卡品造模,方法简便,但死亡率较高,仅15%-20%的动物造模成功。BMSCs源神经干细胞移植后可在海马内存活,向周围的脑区内迁移和整合。结论BMSCs源的神经干细胞在体外可诱导为γ-氨基丁酸能神经元并且对慢性癫痫有一定的治疗作用。Objective To investigate the method of induced differentiation of bone marrow stromal cells （BMSCs） into neuron like cells and the therapeutical effect of their transplantation in animal models of epilepsy. Methods The BMSCs were isolated and purified for 72h in sterile condition, then were cultured in medium DMEM/ F12 which contained bFGF 10 ng/ml and RA 0. 5 p.mol/L. Part of them were used to detect nestin antigen. The remaining part continued induction in DMEM/F12 which contained bFGF 10 ng/ml, RA 0. 5 p.mol/L, NT-3 20 ng/ ml, and BDNF 20 ng/ml, and were stained for GAD 67 after 4 days. Pilocarpine was injected into rats subcutane- ously. Behavioral tests were performed to screen the qualified epilepsy model rats. The neural stem cells originated from BMSCs and neurotrophic factors were transplanted into the hippocampus of epilepsy models with stereoinjection technique. ＂The therapeutical effect of trasplantation was evaluted by frozen section immunohistochemistry and ethology. Results Bone marrow stromal cells were positive for nestin 72h after induction and after 4 days the expression of GAD67 possessed 60%. 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. Conclusion Neural stem cells originated from BMSCs can be induced to differentiated into GABAergic neurons and have some therapeutical effect on epilepsy.

关 键 词：骨髓基质干细胞 Γ-氨基丁酸能神经元 移植 癫痫 大鼠 

分 类 号：R742.1[医药卫生—神经病学与精神病学]