Effect of combined transplantation of neural stem cells and dopaminergic neurons on dyskinesia in rats with Parkinson disease

LI Zhigao; XU Jiaotian; CHEN Xiaoxiang; LIN Hai; SONG Xiaobin; YANG Zhiyong; DENG Xingli

doi:10.3969/j.issn.1002-266X.2017.20.007

Abstract：Objective To explore the efficacy of combined transplantation of neural stem cells (NSCs) and dopaminergic (DA) neurons in treatment of dyskinesia in rats with Parkinson disease (PD).Methods Thirty-six rat models with PD were established by using the classical 6-hydrodopamine damage protocol, and followed by 4-week rotation induction via apomorphine (APO) intraperitoneal injection.These 36 PD rats were randomly divided into the following groups after 4 weeks of APO intraperitoneal injection: the control group, the DA neurons group, the NSCs group, and the NSCs-DA group, and normal saline (NS), CM-DiI labeled DA neurons, GFP-SPIO double labeled NSCs, and CM-Dil labeled DA neurons combined with double labeled NSCs neurons were transplanted into the right striatum of these groups by stereotactic injection, respectively.The rotation frequency of these groups was calculated at week 2, 4, 6, and 8 after the transplantation.Moreover, the NSCs group and DA-NSCs group were scanned by MRI at week 1, 2, 4, and 8.The brain tissues of these groups were collected to observe the migration and homing of the transplanted cells after 8 weeks through prussian blue staining, and the differentiation of NSCs was measured by immunofluorescence.Results The rotation frequencies of the control group, the DA neurons group, the NSCs group, and the NSCs-DA group were decreased gradually, and significant difference was found between every two groups, all P<0.01.The results of MRI scan between NSCs group and NSCs-DA group were as following: over the time of transplantation, the hypointense signal area in T2 W/FFE imaging of both groups were larger than before, and the area in the NSCs-DA group was larger than that of the NSCs group.The results of prussian blue staining suggested that there were a lot of blue particles in the cytoplasm of NSCs and its differentiated cells, which were grafted from the NSCs group and NSCs-DA group.However, just a few cells migrated from the transplantation needle core to the distant, but the most of these blue particles were located on the transplantation sites.And the number of the blue particles in the NSCs-DA group was higher, and its diffusion range was larger than that of the NSCs group.Moreover, there were much more Nestin, TH and GFP positive cells in the NSCs-DA group than in the NSCs group, while the GFAP positive cells and the rates of GFAP/GFP were less than those of the NSCs group (all P<0.01).Conclusion The combined transplantation of NSCs and DA neurons can improve the movement disorder of PD rats significantly, which helps to promote the grafted NSCs homing, migration, and differentiation into DA neurons.