联合培养胚鼠的黑质及纹状体LGE细胞脑内移植治疗帕金森病大鼠的实验研究

目的　观察并检测胚鼠纹状体外侧节突 (LGE)对多巴胺能 (DA)细胞存活性的促进和营养导向作用。方法　将帕金森病 (PD)模型随机分成四组 :Co -culture组 (n =12 ) ;Cograft组 (n =12 ) ;Solo -VM组 (n =12 ) ;Con trol组 (n =8)。将胚鼠LGE细胞和腹侧中脑组织 (VM )制成细胞悬液 ,植入Control组外的其他各组动物的尾壳核。 2周后进行PD鼠行为学检测 ,连续观察 2 4周 ,继之将各组大鼠处死 ,进行免疫组化染色。结果　Co -culture组和Co - graft组大鼠移植后旋转行为较Solo -VM组大鼠明显减少。CO -culture组和CO - graft组之间大鼠的旋转行为比较 ,无统计学差异。免疫组化观察证实LGE和VM离体培养移植和新鲜移植均能提高DA细胞的存活性 ,增加宿主纹状体内DA纤维重新支配的密度 ,并形成明显的DA细胞团。结论　LGE细胞对VM移植物有明显的营养导向作用 ,并可增强DA细胞的存活 ,促进移值后DA细胞功能持久维持 ,并增加DA细胞再支配的密度

Co - cultured embryonic striatum increases the survival of grafted embryonic dopamine neurons

Objective The aim of the present study was to examine the possibility of stimulating fiber outgrowth of grafted neurons by simultaneously grafting dopamine neurons with their embryonic target cells and to characterize further the potential trophic effects of striatal co-grafts and/or co-cultures on grafted mesencephalic DA neuron survival.Method Unilateral 6-hydroxydopamine lesioned Sprague-Dawley(SD)rats were grafted with either a suspension of mesencephalic cells or with both mesencephalic and striatal cell suspensions.Co-grafts were either mixed together or co-cultured.Lesioned rats receiving no graft served as controls.Rotational behavior was assessed following apomorphine challenge at 2 weeks prior to grafting and at 2?4?6 and 8 weeks following grafting 24 weeks post transplantation,all of the rats were sacrificed for HE and Tyrosine hydroxylase immunohistochemistry staining.Results Based on in vitro and in vivo observations of the enhancing effects of striatal tissue on nigral dopaminergic cell development and survival,we demonstrate that inclusion of embryonic striatal cells,specifically from the lateral ganglionic eminence(LGE),produces dopaminergic transplants with augmented functional effects.Rats neonatally DA-depleted and co-transplanted with embryonic nigral and LGE cells developed improved functional outcome when compared with animals receiving only nigral cells(P<0.05),and they required the transplantation of fewer nigral cells to produce a strong behavioral effect.But comparison between the co-culture group and the co-graft group revealed no significant differences(P>0.05).Anatomically,the inclusion of LGE cells produced increased DA cell survival,a higher density of reinnervation into the DA-depleted host striatum,and patches of DA fibers within the co-transplantations.Conclusion These results suggested that nigral-striatal co-grafts,inclusion of in vitro co-cultures and in vivo co-grafts,increased both DA neuron survival and neurite extension fron the mesencephalic component of the grafts.The ability to enhance function,cell survival,reinnervation,and host activation with nigral-striatal co-transplantation in the presence of fewer nigral cells supported the hypothesis of a trophic influence of striatal cells on nigral DA cells.The co-grafted LGE and nigral cells might act as a source of donor of transplantation.