CAJ | 학술논문

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(2 g/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2 μL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5 d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.
배경:배태간세포경유도산생신경전체세포,재진행뇌내이식,의구구비증식여분화적잠능,여숙주신경조직정합,사기유강적가소성,저장유조우파금삼병적치료효과관찰.목적:관찰소서배태간세포분화위신경전체세포,이급정점이식대개진형파금삼병대서모형행위학적영향.설계:수궤대조동물실험.단위:해방군제삼군의대학기출의학부생이학교연실화신경생물학교연실.재료:건강성년Wistar대서50지,수궤분위실험조45지,대조조5지.방법:①실험조대서흑질치밀부여중뇌복측배개구량점각주사6-간기다파5μL(2 g/L)제비파금삼병대서모형,대조조주사생리염수5μL/점,술후1주개시행위학검측,측정모형성공솔,1차/주,련속7주.②선취성공파금삼병모형대서20지실시문상체신경전체세포이식,제량위2μL[세포현액기수위(5～8)×106개/μL].령취5지모형대서문상체주사생리염수2 μL위생리염수대조조.주요관찰지표:①파금삼병모형성공솔.②신경전체세포이식대파금삼병모형대서선전차수적영향.③이식적신경전체세포재체분포,존활여분화정황.결과:①6주후실험조45지대서중공유33지(73%)체도성공파금삼병모형표준.②세균배양명상용무혈청배양기배양형성배태체5 d후,약85%배태간세포분화위Nestin양성적신경전체세포.③신경전체세포뇌내이식후적파금삼병대서적선전차수명현소우생리염수대조조.다수이식적신경전체세포존활,부분분화위TH양성신경원.결론:소서배태간세포유도적신경전체세포뇌내이식후,분화위TH양성신경원,파금삼병대서선전차수명현감소.
BACKGROUND: After neural precursor cells (NPCs) induced from embryonic stem cells (ESCs) have been grafted into the brain, it would still keep some potency of proliferation and differentiation, strong plasticity and integration into the host neural tissues, which would help to observe the therapeutic effect of PD.OBJECTIVE: To observe the differentiation of mouse ESCs into NPCs and the therapeutic effect of NPCs after being transplanted on the behavior of Parkinson disease (PD) rats.DESIGN: Randomly and controlled animal experiment.SETTING: Staff Room of physiology and Staff Room of Neurobiology, Depayment of Basic Medical Sciences, the Third Military Medical University of Chinese PLA.MATERIALS: Totally 50 healthy adult Wistar rats were chosen and randomly divided into experimental group (n=45) and control group (n=5).METHODS: ① 5 μ L (2 g/L)6-hydroxydopamine (6-OHDA) was injected into substantia nigra pars compacta and ventral tegmental area two points in the experimental group to prepare PD rats, and normal saline with the dosage of 5 μL per point was injected into the rats in the control group.Behavioral test began at 1 week after operation to measure successful rate of model establishing, once a week for 7 consecutive weeks. ② Totally 20 successful PD rat models were chosen to perform corpus striatum NPCs with the dosage of 2 μL [the count of cell suspension was (5-8)×106/μL].The other 5 rats were given 2 μL normal saline at corpus striatum as normal saline control group.MAIN OUTCOME MEASURES: ① Successful rate of PD model. ②Effect of NPCs transplantation on the rotation times of PD models. ③ Distribution of transplanted NPCs in vivo, and survival and differentiation.RESULTS: ①6 weeks later, totally 33 of 45 rats in the experimental group achieve the standard of PD model . ② About 85% of mouse ESCs were differentiated into Nestin-positive NPCs 5 days after the embryoid bodies formed in the bacterial dishes and cultured in the N2 serum-free medium. ③The rotation times of the PD rats was significantly decreased after the intracerebral transplantation of NPCs as compared with normal control group. Most of the NPCs grafted into striatum of PD rats were survived, and some were differentiated into TH-positive neurons.CONCLUSION: The mouse ESCs-derived NPCs could be transplanted into striatum of PD rats, and then differentiated into TH-positive neurons,which leads to the obvious decrease of rotation times.