帕金森病模型大鼠神经干细胞移植后的迁移

Migration of Neural Stem Cells after Transplanted in Parkinsonian Rat

首先分离培养E14胚胎SD(Sprague-Dawley)大鼠脑室下带区(subventricular zone,SVZ)神经干细胞,用携带增强型绿色荧光蛋白(enhanced green fluorescent protein,EGFP)基因的2型重组腺相关病毒(rAAV2)进行感染,获得具有EGFP标记的神经干细胞。运用单侧损毁成年SD大鼠黑质多巴胺能神经元的帕金森病(Parkinson’sdisease,PD)动物模型,将EGFP标记后的细胞分别移植到PD模型动物的纹状体或黑质内。在移植后120d取移植大鼠全脑进行矢状和冠状连续冰冻切片观察细胞的迁移行为,切片厚度30μm。结果显示,移植位点始终能检测到标记细胞,移植到黑质的细胞在黑质部位迁移扩散,移植到纹状体的细胞成条链式排列沿腹后内侧方向迁移到达黑质,细胞迁移表现出明显的方向性。实验结果提示移植的神经干细胞具有向损伤的黑质区域迁移并长期稳定在黑质存活的特性。

The neural stem cells （NSCs） from E14 rat subventricular zone （SVZ） were isolated and amplified as the cell source for transplantation. The cells were labeled by enhanced green fluorescent protein （EGFP） using recombinant adeno-associated virus （rAAV2） containing the marker gene of EGFP. The adult Sprague-Dawley rats were used to establish Parkinson＇s disease （PD） models by unilateral microinjection of 6-hydroxydopamine （6- OHDA） which damaged dopaminergic neurons in the substantia nigra. EGFP labeled NSCs were transplanted into substantia nigra pars compacta （SNc） or striatum of PD rat models. To observe the behavior of transplanted NSCs, rats were sacrificed 120 days after transplantation and brains were cryosectioned serially in the coronal and saggital plane at the thickness of 30 μm. The results showed that EGFP labeled cells were seen in injection sites of all rats. The cells transplanted into SNc migrated and dispersed in SNc. The cells transplanted into striatum migrated ventrally, posterior and internally to SNc in chains and showed SNc special direction significantly. This phenomenon implicated that the transplanted NSCs have the characteristic of migration to SNc and could survive at SNc for a long time.