Aleksandrova M A, Poltavtseva R A, Revishchin A V, Korochkin L I, Sukhikh G T
Laboratory of Experimental Neurobiology, Institute of Developmental Biology, RAS, Moscow.
Morfologiia. 2003;123(3):17-20.
The purpose of the present study was the investigation of human neural stem/progenitor cells (SPC) cultured in vitro, with special reference to their capacity for grafting, migration and differentiation after transplantation into adult rat brain. SPC were isolated from the brain of 9-week-old human embryos and were cultured in a selective medium for 3 weeks. For transplantation, cell suspension or whole neurospheres were used; they were studied 4 weeks following the transplantation in hippocampus, striatum and lateral ventricle of adult rat brain. For the analysis of transplanted SPC, various histological and immunohistochemical staining methods were applied (bisbenzidine, BrdU, antibodies against human nuclei, vimentin, beta-tubulin, neurofilaments, GFAP), that allowed an independent evaluation of their state and differentiation. Transplanted human brain SPC were shown to survive well for one month in all the areas of adult rat brain without immunosuppression. Cells from suspension transplants actively migrated and differentiated into neurons and glial cells. Meanwhile, cell migration from the transplanted whole neurospheres was limited or absent due to the formation of glial barrier.
本研究的目的是对体外培养的人神经干细胞/祖细胞(SPC)进行研究,特别关注其移植到成年大鼠脑内后的移植、迁移及分化能力。SPC从9周龄人胚胎的脑中分离出来,并在选择性培养基中培养3周。移植时使用细胞悬液或完整的神经球;在将其移植到成年大鼠脑的海马体、纹状体和侧脑室4周后对其进行研究。为了分析移植的SPC,应用了各种组织学和免疫组织化学染色方法(双苯并咪唑、BrdU、抗人核抗体、波形蛋白、β-微管蛋白、神经丝、GFAP),从而能够独立评估它们的状态和分化情况。结果显示,移植的人脑SPC在成年大鼠脑的所有区域中,在未进行免疫抑制的情况下能良好存活一个月。悬浮移植的细胞能积极迁移并分化为神经元和神经胶质细胞。同时,由于胶质屏障的形成,来自移植完整神经球的细胞迁移受到限制或不存在。