中国组织工程研究

中国组织工程研究 ›› 2016, Vol. 20 ›› Issue (45): 6733-6738.doi: 10.3969/j.issn.2095-4344.2016.45.007

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

还原型谷胱苷肽对骨髓基质干细胞增殖及向神经样细胞分化的影响

冯林杰，甘洪全，余向前，杨  健，白卫飞，刘英杰   

  1. 华北理工大学附属医院骨科，河北省唐山市  063000
  • 修回日期:2016-08-29 出版日期:2016-11-04 发布日期:2016-11-04
  • 通讯作者: 刘英杰，硕士，教授，华北理工大学附属医院骨科，河北省唐山市 063000
  • 作者简介:冯林杰，男，1980年生，河北省邯郸市人，汉族，2008年第四军医大学毕业，硕士，主治医师，主要从事脊柱外科方面的研究。
  • 基金资助:

    河北省医学科学研究课题计划项目(20150508)

Reduced glutathione effect on the proliferation and neuronal differentiation of bone marrow stromal stem cells

Feng Lin-jie, Gan Hong-quan, Yu Xiang-qian, Yang Jian, Bai Wei-fei, Liu Ying-jie   

  1. Department of Orthopaedics, North China University of Science and Technology Affiliated Hospital, Tangshan 063000, Hebei Province, China
  • Revised:2016-08-29 Online:2016-11-04 Published:2016-11-04
  • Contact: Liu Ying-jie, Master, Professor, Department of Orthopaedics, North China University of Science and Technology Affiliated Hospital, Tangshan 063000, Hebei Province, China
  • About author:Feng Lin-jie, Master, Attending physician, Department of Orthopaedics, North China University of Science and Technology Affiliated Hospital, Tangshan 063000, Hebei Province, China
  • Supported by:

    the Medical Science Research Project of Hebei Province, No. 20150508

PDF

179

摘要：

文章快速阅读：

文题释义：
还原型谷胱甘肽：是自然界广泛存在的三肽类物质，主要存在于细胞质中，在多种细胞生化功能中起作用。还原型谷胱甘肽中半胱氨酸的巯基是高度亲核的，因此它是异生物或者它们的代谢物激发的亲电物的主要靶结构，从而对主要亲核位点起保护作用，而后者如果受影响会导致细胞损伤。
诱导分化为神经样细胞的方法：常用的诱导方法有诱导剂诱导法、微环境诱导法和基因定向诱导分化法等。诱导剂诱导分化法常用的诱导剂包括神经营养因子和细胞因子，如碱性成纤维细胞生长因子、乙酰胆碱等，化学试剂包括抗氧化剂如β-巯基乙醇等，局部微环境调控定向分化法包括与神经组织或细胞共培养，如嗅鞘细胞等，另外就是基因定向诱导的方法，不同转录因子的表达导致不同谱系的分化。目前应用基因定向诱导的方法是未来研究的方向和趋势。

 

摘要
背景：骨髓基质干细胞具有易获取及多向分化潜能，通过诱导骨髓基质干细胞分化获得神经干细胞是治疗脊髓损伤的重要途径和方法。
目的：观察不同浓度还原型谷胱苷肽对骨髓基质干细胞增殖及向神经样细胞分化的影响。
方法：配置5组不同浓度的还原型谷胱苷肽(0，5，10，20，40 mmol/L)培养液，干预骨髓基质干细胞72 h后，采用MTT法分析细胞增殖活性，倒置显微镜下连续观察细胞形态变化，免疫组织荧光染色法测定MAP-2和GFAP表达。
结果与结论：①低浓度5 mmol/L组还原型谷胱苷肽培养液能够促进骨髓基质干细胞增殖(P < 0.05)，20 mmol/L组和40 mmol/L组还原型谷胱苷肽培养液抑制细胞增殖(P < 0.05)，10 mmol/L还原型谷胱苷肽组细胞集落的数量和大小与对照组未见明显差别；②10 mmol/L组还原型谷胱苷肽培养液诱导骨髓基质干细胞向神经细胞分化作用较明显，荧光显微镜下可见大部分细胞表达MAP-2和GFAP，随机选择5个视野观察并计算骨髓基质干细胞分化为神经样细胞的比率为(62.0±4.8)%；③结果证实，低浓度 (5 mmol/L)还原型谷胱苷肽可以促进骨髓基质干细胞增殖，10 mmol/L还原型谷胱苷肽有较强的诱导骨髓基质干细胞向神经样细胞分化的作用。

 

 

关键词: 干细胞, 骨髓干细胞, 还原型谷胱苷肽, 骨髓基质干细胞, 诱导分化, 神经细胞

Abstract:

BACKGROUND: As bone marrow stromal stem cells can be easily obtained and have multi-directional differentiation potential, it is an important approach to obtain neural stem cells for treatment of spinal cord injury through induced differentiation of bone marrow stromal stem cells.
OBJECTIVE: To observe the effects of different concentrations of reduced glutathione on proliferation and neuronal differentiation of bone marrow stromal stem cells.
METHODS: Bone marrow stromal cells were isolated and cultured by limited dilution method. The cloned bone marrow stromal cells were stimulated with reduced glutathione at different concentrations (0, 5, 10, 20, 40 mmol/L) respectively. After 72 hours of culture, proliferation of bone marrow stromal cells was examined by MTT assay. The morphology of bone marrow stromal cells was observed under inverted microscope and the cells were identified by immunofluorescence staining of microtubule associated protein-2 and glial fibrillary acidic protein.
RESULTS AND CONCLUSION: We found that 5 mmol/L reduced glutathione medium could promote the proliferation of bone marrow stromal cells (P < 0.05), but reduced glutathione at 20 and 40 mmol/L inhibited the cell proliferation (P < 0.05). The number and size of cell colonies in the 10 mmol/L reduced glutathione group had no obvious difference from the control group. Reduced glutathione at 10 mmol/L was powerful to induce the neuronal differentiation of bone marrow stromal stem cells, and most cells expressed microtubule associated protein-2 and glial fibrillary acidic protein. A random selection of 5 fields of view was conducted and percentage of bone marrow stromal cells differentiating into neuron-like cells was calculated as (62.0±4.8)%. These results confirmed that low-concentration (5 mmol/L) reduced glutathione can promote the proliferation of bone marrow stromal cells, while 10 mmol/L reduced glutathione has a strong induction of bone marrow stromal cells differentiating into neuron-like cells.

 

 

Key words: Bone Marrow, Mesenchymal Stem Cells, Protein Disulfide Reductase (Glutathione), Cell Differentiation, Neurons, Tissue Engineering

中图分类号: