中国组织工程研究

中国组织工程研究 ›› 2018, Vol. 22 ›› Issue (13): 2039-2044.doi: 10.3969/j.issn.2095-4344.0490

• 脐带脐血干细胞 umbilical cord blood stem cells • 上一篇    下一篇

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

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   

  1. 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
  • 修回日期:2017-12-24 出版日期:2018-05-08 发布日期:2018-05-08
  • 通讯作者: 赵宗茂，博士，教授，博士生导师，河北医科大学附属第二医院， 河北省石家庄市 050000
  • 作者简介:张鹏，男，1980 年生，河北省邯郸市人，汉族，2006年河北医科大学毕业，硕士，主治医师，主要从事神经外科专业相关研究。

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

Zhang Peng1, Zhao Zong-mao2, Li Jian-hua1, Liu Hui1, Liu Yong-jun1, Li Min-jie1, Chen Ming-wei1, Shen Lun1,He Lei1   

  1. 1the First Hospital of Handan, Handan 056000, Hebei Province, China; 2the Second Hospital of Hebei Medical University, Shijiazhuang 050000, Hebei Province, China
  • Revised:2017-12-24 Online:2018-05-08 Published:2018-05-08
  • Contact: Zhao Zong-mao, M.D., Professor, Doctoral supervisor, the Second Hospital of Hebei Medical University, Shijiazhuang 050000, Hebei Province, China
  • About author:Zhang Peng, Master, Attending physician, the First Hospital of Handan, Handan 056000, Hebei Province, China

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摘要：

文章快速阅读：

文题释义：
单唾液酸四己糖神经节苷脂：是从哺乳动物神经组织中提取的一种物质，可以穿透血脑屏障，减轻脑组织水肿，清除自由基，促进神经细胞的再生以及轴突和突触的生长，以达到促进中枢神经系统损伤修复的作用。临床已广泛应用于各种原因所致神经病变，并已取得良好的效果，是临床常用的脑神经保护剂之一。
间充质干细胞体外向神经元样细胞分化的机制：目前尚未形成定论，有些观点认为细胞发生神经元样变是一系列的基因表达事件，而另外的观点则认为间充质干细胞的形态变化仅仅是因为细胞骨架破坏导致胞浆收缩形成的。大多数诱导而来的神经元样细胞在体外存活的时间很短，且神经元样细胞有无神经电生理活动还有待于进一步验证。

 

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。

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程
ORCID:0000-0002-9805-2743(张鹏)

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

Abstract:

BACKGROUND: Studies have confirmed that monosialotetrahexosyl anglioside 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. 

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Umbilical Cord, Mesenchymal Stem Cells, G(M1) Ganglioside, Cell Differentiation, Neurons, Tissue Engineering

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