CAJ | 학술논문

目的研究脂肪来源的间充质干细胞诱导为γ-氨基丁酸(GABA)能神经元的方法 ,探讨GABA能神经元移植治疗帕金森病模型大鼠的疗效. 方法取大鼠脂肪组织.利用本单位自行配制的神经干细胞培养基诱导分化为神经干细胞,利用GABA能神经元定向诱导培养基对神经干细胞进行二次定向诱导,并对其进行特异性鉴定.将诱导成功的神经干细胞、GABA能神经元分别移植入帕金森病大鼠模型的丘脑底核,在移植后2周、4周、8周观察大鼠行为学变化情况.结果体外扩增的脂肪间充质干细胞经过神经干细胞培养基培养后,细胞定向诱导并表达巢蛋白、神经元烯醇化酶(NSE)等神经干细胞标志.经GABA能神经元定向分化培养基二次诱导后,免疫荧光鉴定细胞GAD65阳性.立体定向移植细胞4周后,神经干细胞组与GABA能神经元组的大鼠行为学均得到改善,且GABA能神经元组的疗效更加显著. 结论脂防来源的间充质干细胞可诱导分化为GABA能神经元,将其移植人大鼠的丘脑底核可以明显改善帕金森病大鼠的运动功能.
목적 연구지방래원적간충질간세포유도위γ-안기정산(GABA)능신경원적방법 ,탐토GABA능신경원이식치료파금삼병모형대서적료효. 방법 취대서지방조직.이용본단위자행배제적신경간세포배양기유도분화위신경간세포,이용GABA능신경원정향유도배양기대신경간세포진행이차정향유도,병대기진행특이성감정.장유도성공적신경간세포、GABA능신경원분별이식입파금삼병대서모형적구뇌저핵,재이식후2주、4주、8주관찰대서행위학변화정황.결과 체외확증적지방간충질간세포경과신경간세포배양기배양후,세포정향유도병표체소단백、신경원희순화매(NSE)등신경간세포표지.경GABA능신경원정향분화배양기이차유도후,면역형광감정세포GAD65양성.입체정향이식세포4주후,신경간세포조여GABA능신경원조적대서행위학균득도개선,차GABA능신경원조적료효경가현저. 결론 지방래원적간충질간세포가유도분화위GABA능신경원,장기이식인대서적구뇌저핵가이명현개선파금삼병대서적운동공능.
Objective To investigate the method for inducing OABAergic neurons from adipose-derived mesenchymal stem cells (ADSCs) and observe the effect of transplantation of these neurons in the treatment of parkinsonian rats. Methods ADSCs isolated from rat inguinal fat pads were digested with collagenase, cultured and passaged in vitro, from which neural stem cells were induced using the neural stem cell culture medium prepared by our institute and identified for the stem cell markers. The neural stem cells obtained were further induced using the GABAergic neuron culture medium. After identification for the marker GAD65, the GABAergic neurons or the neural stem cells were stereotaxically transplanted into the subthalamic nucleus of the Parkinsonian rats, and the behavioral changes of the rats were observed at 2, 4 and 8 weeks after the cell transplantation. Results The neural stem cells differentiated from the ADSCs expressed the stem cell markers including nestin and neuron-specific enolase. After the second induction, the cells were positive for GAD65 as identified by immunofluorescence staining. Four weeks after transplantation of the neural stem cells and GABAergic neurons into the subthalamic nucleus, the parkinsonian rats exhibited significantly improved rotational behavior induced by apomorphine, and the improvement was especially obvious in rats with GABAergic cell transplantation. Conclusion GABAergic neurons can be induced from the rat ADSCs and transplantation of these neurons into the subthalamic nucleus can produce obvious behavioral improvement in rat models of Parkinson disease.