食蟹猴多功能干细胞向运动神经元定向分化  

Pluripotent Stem Cells Differentiate into Motor Neurons in Cynomolgus Monkeys

作　　者：曹静 李梦佳 王芳 曾玉强 魏景宽 牛昱宇[1] CAO Jing;LI Mengjia;WANG Fang;ZENG Yuqiang;WEI Jingkuan;NIU Yuyu(Kunming University of Science and Technology,Kunming 50500,China)

机构地区：[1]昆明理工大学生命科学与技术学院

出　　处：《中国细胞生物学学报》2019年第10期1976-1985,共10页Chinese Journal of Cell Biology

基　　金：国家重点研发计划干细胞专项(批准号:2018YFA0108503);云南省基础研究计划面上项目(批准号:2018FB118)资助的课题~~

摘　　要：运动神经元(motor neuron,MN)是位于大脑皮层、脊髓前角、脑干等区域的一种终末分化的细胞,MN的损伤涉及多种神经退行性疾病。目前,体内研究该类疾病面临巨大困难。因此利用体外分化手段获得成熟的MN,对于研究神经退行性疾病的致病机制和治疗具有重大意义。非人灵长类与人具有高度相似的遗传背景,作为一种理想动物模型已经被广泛应用于疾病的研究中。因此,该研究利用食蟹猴(Macaca fascicularis)的诱导多功能干细胞(induced pluripotent stem cells,iPSCs),建立了成熟MN快速且高效的分化体系。该实验的分化体系共计28天,且包括四个分化阶段,最终90%以上的MN均处于成熟状态。四个分化阶段为:(1)iPSCs向神经上皮细胞(neuroepithelial progenitors,NEP)的分化;(2)NEP向运动神经元前体细胞(motor neuron progenitors,MNP)的分化;(3)MNP向非成熟MN的分化;(4)非成熟的MN逐渐成熟。该研究首次成功建立食蟹猴iPSCs到MN的分化体系,并确定多聚-L-鸟氨酸和层黏连蛋白(lamina-521)双重包被的培养皿、细胞的接种密度以及ROCK抑制剂的添加可提高MN的存活率,为今后探究运动神经元疾病的发病机制和干细胞移植治疗奠定了良好的基础。MN(motor neuron)is a specialized type of neuron whose cell body is located in the motor cortex,brainstem,and the spinal cord.The damage of this type of cell results in many neurodegenerative diseases.So far,it is extremely difficult to study these diseases in vivo.Therefore,it is of great significance to study the pathogenesis and therapy of neurodegenerative diseases through the differentiation of mature MN in vitro.Cynomolgus monkeys(Macaca fascicularis),as a non-human primate model,has a highly similar genetic background with humans and is widely used for the study of these diseases.We developed a method to guide the induced pluripotent stem cell of cynomolgus monkeys into an enriched population(>90%)of mature MNs in 28 days,which consists of four differentiation stages.The four stages are as follows:(1)iPSCs(induced pluripotent stem cells)were induced into NEP(neuroepithelial progenitors);(2)NEP were induced into MNP(motor neuron progenitors);(3)MNP were induced into non-mature MN;(4)non-mature MN were induced into mature MN.This study successfully established a method for guiding iPSCs of cynomolgus monkeys into MN.Moreover,it showed that the poly-L-ornithine and lamina-521 double-coated dish,cell density,and the addition of ROCK inhibitor(Y27632)can contribute to the survival of MN.In summary,this study provides a basis for studying the pathogenesis and stem cell transplantation therapy of MN degenerative diseases.

关 键 词：诱导多能干细胞 神经干细胞 运动神经元 神经退行性疾病 

分 类 号：R741[医药卫生—神经病学与精神病学]