同种异体神经干细胞脑内移植免疫排斥反应的实验研究

目的观察帕金森病(PD)模型大鼠同种异体神经干细胞(NSC)脑内移植是否存在免疫排斥反应。方法取E14.5d SD胚鼠腹侧中脑组织进行分离、培养、扩增,经Brdu、Nestin免疫组化染色鉴定确认为NSC后,借助脑立体定位仪移植到SD大鼠PD模型的纹状体内,分别于10、21、35、60d时检测其旋转行为后分批处死,再行HE、CD4、CD8、酪氨酸羟化酶(TH)和MHC-Ⅱ抗原免疫组化染色。结果移植组10d时,CD4、CD8、、MHC-Ⅱ少量表达,以21d时较为明显,35d时明显减少,60d消失;10d、21d时移植区未见明显TH阳性神经元,35d后数量显著增加。阳性对照组在10d、21d时见少量CD4、CD8、和MHC-Ⅱ阳性细胞,35d时消失,各时间点都未见TH阳性神经元。阴性对照组在各时间点都未见TH阳性神经元和CD4、CD8、MHC-Ⅱ阳性细胞。PD模型移植组旋转行为35d时出现改善。结论神经干细胞脑内移植未见明显免疫排斥反应。

Experimental research on immunological rejection in neural stem cells allograft

AIM: To investigate the occurrence of immunological rejection in brain transplantation of neural stem cells (NSCs) in the rat of Parkinson's disease model. METHODS: NSCs derived from the brain of E14.5d SD rat were cultured in vitro. Single cell suspensions were grafted into the striatum of the rat model of Parkinson's disease. Surviving animals were sacrificed at 10, 21, 35 and 60 d after transplantation, and the brain tissue was stained with HE and immunocytochemically to detect the expression of tyrosine hydroxylase (TH), CD4, CD8 and major histocompatibility complex class II antigens (MHC-II). RESULTS: The expression of elicited CD4, CD8 and MHC-II was detected within and around the allografts in the graft groups at 10 and 21 d after grafting and was subsiding at 35 d. At 60 d only very occasional immunopositive cells were present. The number of TH positive neurons was low at 10, 21 d, but increased at 35 d and 60 d. There was no significant difference between negative and positive control groups at different time points. The rotation behavior of PD was decreased 35 d after transplantation. CONCLUSION: Intracerebral transplantation of NSCs did not cause remarkable immunological rejection.