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背景:胚胎干细胞经诱导产生神经前体细胞,再进行脑内移植,依旧具备增殖与分化的潜能,与宿主神经组织整合,使其有强的可塑性,这将有助于帕金森病的治疗效果观察。目的:观察小鼠胚胎干细胞分化为神经前体细胞,以及定点移植对改进型帕金森病大鼠模型行为学的影响。设计:随机对照动物实验。单位:解放军第三军医大学基础医学部生理学教研室和神经生物学教研室。材料:健康成年Wistar大鼠50只,随机分为实验组45只,对照组5只。方法:①实验组大鼠黑质致密部与中脑腹侧背盖区两点各注射6-羟基多巴5μL(2g/L)制备帕金森病大鼠模型,对照组注射生理盐水5μL/点,术后1周开始行为学检测,测定模型成功率,1次/周,连续7周。②选取成功帕金森病模型大鼠20只实施纹状体神经前体细胞移植,剂量为2μL[细胞悬液记数为(5～8)×106个/μL]。另取5只模型大鼠纹状体注射生理盐水2μL为生理盐水对照组。主要观察指标:①帕金森病模型成功率。②神经前体细胞移植对帕金森病模型大鼠旋转次数的影响。③移植的神经前体细胞在体分布,存活与分化情况。结果:①6周后实验组45只大鼠中共有33只(73%)达到成功帕金森病模型标准。②细菌培养皿上用无血清培养基培养形成胚胎体5d后,约85%胚胎干细胞分化为Nestin阳性的神经前体细胞。③神经前体细胞脑内移植后的帕金森病大鼠的旋转次数明显少于生理盐水对照组。多数移植的神经前体细胞存活,部分分化为TH阳性神经元。结论:小鼠胚胎干细胞诱导的神经前体细胞脑内移植后,分化为TH阳性神经元,帕金森病大鼠旋转次数明显减少。
BACKGROUND: Embryonic stem cells are induced to produce neural progenitor cells and then undergo intracerebral transplantation. They still have the potential to proliferate and differentiate and integrate with the host nerve tissue to have strong plasticity, which will be helpful for the treatment of Parkinson’s disease Observed. OBJECTIVE: To observe the differentiation of mouse embryonic stem cells into neural precursor cells and the effect of site-directed transplantation on the behavior of model rats with modified Parkinson’s disease. Design: Randomized controlled animal experiments. Unit: Department of Physiology and Neurobiology, Department of Basic Medicine, Third Military Medical University. MATERIALS: Fifty healthy adult Wistar rats were randomly divided into experimental group (n = 45) and control group (n = 5). Methods: ① The rat model of Parkinson’s disease was prepared by injecting 5μL of 6-hydroxydopamine (2g / L) into both substantia nigra pars compacta and ventral tegmental area in experimental group. The control group was injected with 5μL / One week after operation, behavioral testing was started. The success rate of the model was measured once a week for 7 weeks. ② Twenty successful rats with Parkinson’s disease were selected for transplantation of striatal neural progenitor cells in a dose of 2 μL [cell suspension count (5 ~ 8) × 106 cells / μL]. Another 5 rats were injected with normal saline striatum 2μL normal saline control group. MAIN OUTCOME MEASURES: ① Parkinson’s disease model success rate. Effect of neural progenitor cell transplantation on the number of rotations in rats with Parkinson’s disease. ③ transplantation of neural precursor cells in vivo distribution, survival and differentiation. Results: ① Six weeks later, 33 out of 45 rats (73%) in the experimental group reached the standard of successful Parkinson’s disease model. (2) About 85% of embryonic stem cells differentiated into Nestin-positive neural progenitor cells cultured in serum-free medium for 5 days on bacterial culture dishes. (3) The number of rotations of Parkinson’s disease rats after transplantation of neural precursor cells in brain was significantly less than that of saline control group. Most of the transplanted neural precursor cells survive and partially differentiate into TH-positive neurons. CONCLUSION: The neural precursor cells induced by mouse embryonic stem cells differentiate into TH-positive neurons after transplantation into the brain. The number of rotations in Parkinson’s disease rats is significantly reduced.