单唾液酸四己糖神经节苷脂最佳质量浓度诱导人脐带间充质干细胞向神经元样细胞的分化  

Monosialotetrahexosyl ganglioside at an optimal concentration: inducing neuron-like differentiation of human umbilical cord mesenchymal stem cells

作　　者：张鹏[1] 赵宗茂[2] 李建华[1] 刘辉[1] 刘永军[1] 李敏捷[1] 陈明伟[1] 申仑[1] 何磊[1] Zhang Peng;Zhao Zong-mao;Li Jian-hua;Liu Hui;Liu Yong-jun;Li Min-jie;Chen Ming-wei;Shen Lun;He Lei(the First Hospital of Handan, Handan 056000, Hebei Province, China;the Second Hospital of Hebei Medical University, Shijiazhuang 050000, Hebei Province, China)

机构地区：[1]邯郸市第一医院,河北省邯郸市056000 [2]河北医科大学附属第二医院,河北省石家庄市050000

出　　处：《中国组织工程研究》2018年第13期2039-2044,共6页Chinese Journal of Tissue Engineering Research

摘　　要：背景:国内外研究证实单唾液酸四己糖神经节苷脂在体外能够诱导人脐带间充质干细胞分化为神经元样细胞,但探索其最合适的诱导剂量等相关研究甚少。目的:探索单唾液酸四己糖神经节苷脂在体外诱导人脐带间充质干细胞分化为神经元样细胞的最佳质量浓度。方法:采用胶原酶消化法获取人脐带间充质干细胞,扩增后,取第3代细胞,随机分为5组,当细胞融合至70%-80%时,分别加入质量浓度为50,100,150,200 mg/L单唾液酸四己糖神经节苷脂的诱导液,空白对照组只加入等量L-DMEM培养基。倒置显微镜下观察细胞形态变化。诱导第6小时用免疫细胞化学染色方法检测GFAP、NF-H、MAP-2的表达。结果与结论:从脐带组织中分离出来的人脐带间充质干细胞可以稳定传代,并表达间充质干细胞表面标志,单唾液酸四己糖神经节苷脂可以诱导人脐带间充质干细胞向神经元样细胞分化,并表达成熟神经细胞标志物MAP-2、NF-H,最佳诱导质量浓度为150 mg/L。BACKGROUND： Studies have confirmed that monosialotetrahexosyl ganglioside can induce human umbilical cord mesenchymal stem cells to differentiate into neuron-like cells, but little is reported on its optimal concentration. OBJECTIVE： To explore the optimal concentration of monosialotetrahexosyl ganglioside that induces human umbilical cord mesenchymal stem cells to differentiate into neuron-like cells in vitro. METHODS： Human umbilical cord mesenchymal stem cells were isolated by using collagenase digestion method, and after expansion, passage 3 cells were randomly allocated into five groups. When 70%-80% of cells were confluent, 50, 100, 150 and 200 mg/L monosialotetrahexosyl ganglioside induction solutions were added in corresponding experimental groups, while cells in the blank control group were cultured in the same volume of L-DMEM medium. Cell morphology was observed under inverted phase contrast microscope. Expression levels of microtubule-associated protein 2, neurofilament protein and glial fibrillary acidic protein were measured by using immunohistochemistry at 6 hours after induction. RESULTS AND CONCLUSION： Human umbilical cord mesenchymal stem cells were isolated successfully and sub-cultured stably. These cells could express surface markers of mesenchymal stem cells. Monosialotetrahexosyl ganglioside at the optimal concentration of 150 mg/L was confirmed to induce the neuron-like differentiation of human umbilical cord mesenchymal stem cells, and differentiated cells could express microtubule-associated protein 2 and neurofilament protein as neuron-specific markers.

关 键 词：干细胞 单唾液酸四己糖神经节苷脂 人脐带间充质干细胞 诱导分化 神经元样细胞 

分 类 号：R394.2[医药卫生—医学遗传学]