中国神经再生研究(英文版) ›› 2015, Vol. 10 ›› Issue (8): 1312-1316.doi: 10.4103/1673-5374.162768

• 原著:神经损伤修复保护与再生 • 上一篇    下一篇

脐带间充质干细胞在海藻酸盐支架中分化为运动神经元样细胞

  

  • 收稿日期:2015-05-14 出版日期:2015-08-24 发布日期:2015-08-24

Differentiation of Wharton’s jelly mesenchymal stem cells into neurons in alginate scaffold

Seyed Mojtaba Hosseini1, 2, 3, Attiyeh Vasaghi1, 2, Newsha Nakhlparvar1, 2, Reza Roshanravan4, Tahereh Talaei-khozani5, 6, *, Zahra Razi2, 7   

  1. 1 Student Research Committee, Shiraz University of Medical Sciences, Shiraz, Iran
    2 Cell and Molecular Medicine Student Research Group, Medical School, Shiraz University of Medical Sciences, Shiraz, Iran
    3 Stem Cell Laboratory, Department of Anatomy, Shiraz University of Medical Sciences, Shiraz, Iran
    4 Colorectal Research Center, Department of Surgery, Shiraz University of Medical Science, Shiraz, Iran
    5 Tissue Engineering Laboratory, Department of Tissue Engineering, School of Advanced Medical Science and Technology, Shiraz University of Medical Sciences, Shiraz, Iran
    6 Laboratory for Stem Cell Research, Department of Anatomy, Medical School, Shiraz University of Medical Sciences, Shiraz, Iran
    7 Department of Medical Physics, Shiraz University of Medical Sciences, Shiraz, Iran
  • Received:2015-05-14 Online:2015-08-24 Published:2015-08-24
  • Contact: Tahereh Talaei-khozani,talaeit@sums.ac.ir.

摘要:

海藻酸盐支架是一种可促进胚胎干细胞向神经元样细胞分化的合适生物材料。由此,我们假设海藻酸盐支架适于培养脐带间充质干细胞,增强其向神经元样细胞分化的能力。实验将脐带间充质干细胞培养于包含0.25%海藻酸盐的3D支架、50mM CaCl2的神经细胞培养基中,并与单纯培养于2D培养条件下的神经细胞培养基中的干细胞进行比较。10d后,应用免疫荧光染色检测到3D海藻酸盐支架培养的脐带间充质干细胞神经元β-tubulin和运动神经元标志物CD 271的表达量明显高于2D培养基。说明3D海藻酸盐支架培养系统可有效诱导脐带间充质干细胞的神经分化。

关键词: 神经再生, 脐带间充质干细胞, 间充质干细胞, 神经元, 运动神经元, 海藻酸盐, 3D支架

Abstract:

Alginate scaffold has been considered as an appropriate biomaterial for promoting the differentiation of embryonic stem cells toward neuronal cell lineage. We hypothesized that alginate scaffold is suitable for culturing Wharton’s jelly mesenchymal stem cells (WJMSCs) and can promote the differentiation of WJMSCs into neuron-like cells. In this study, we cultured WJMSCs in a three-dimensional scaffold fabricated by 0.25% alginate and 50 mM CaCl2 in the presence of neurogenic medium containing 10 μM retinoic acid and 20 ng/mL basic fibroblast growth factor. These cells were also cultured in conventional two-dimensional culture condition in the presence of neurogenic medium as controls. After 10 days, immunofluorescence staining was performed for detecting β-tubulin (marker for WJMSCs-differentiated neuron) and CD271 (motor neuron marker). β-Tubulin and CD271 expression levels were significantly greater in the WJMSCs cultured in the three-dimensional alginate scaffold than in the conventional two-dimensional culture condition. These findings suggest that three-dimensional alginate scaffold cell culture system can induce neuronal differentiation of WJMSCs effectively.

Key words: nerve regeneration, Wharton’s jelly mesenchymal stem cells, mesenchymal stem cells, neurons, motor neurons, alginate, 3D scaffold, neural regeneration