基础医学与临床 ›› 2019, Vol. 39 ›› Issue (5): 630-635.

• 研究论文 • 上一篇    下一篇

优化海藻酸钠-壳聚糖多孔复合支架用于人脐带来源间充质干细胞的体外扩增

刘雨杰1,赵锦1,何志旭2,曾令文1   

  1. 1. 贵州医科大学
    2. 贵阳医学院组织工程与干细胞中心
  • 收稿日期:2018-06-25 修回日期:2018-09-25 出版日期:2019-05-05 发布日期:2019-04-24
  • 通讯作者: 曾令文 E-mail:zeng_lingwen@gibh.ac.cn
  • 基金资助:
    国家重大科学研究计划

Optimization of alginate-chitosan porous composite scaffold for in vitro proliferation of human umbilical cord-derived mesenchymal stem cells

  • Received:2018-06-25 Revised:2018-09-25 Online:2019-05-05 Published:2019-04-24

摘要: 目的 通过制备及优化海藻酸钠-壳聚糖多孔复合支架用于人脐带来源的间充质干细胞体外扩增,为干细胞在组织工程中的应用与疾病治疗提供实验基础。方法 通过制备海藻酸钠-壳聚糖多孔复合支架,并接枝多聚赖氨酸对其优化。然后,从产妇脐带中分离出间充质干细胞,接种在复合支架表面,监控其增殖情况。利用细胞计数法和CCK-8试剂盒,绘制细胞增殖曲线,并用β-半乳糖苷酶衰老检测试剂盒评估干细胞的衰老程度。结果 成功制备并优化了海藻酸钠-壳聚糖复合支架,发现在接枝多聚赖氨酸的支架上接种间充质干细胞,细胞能够很好地附着,覆盖在复合支架表面,且能快速增殖。结论 优化的海藻酸钠-壳聚糖多孔复合支架能为细胞增殖提供优良环境,具备体外扩增间充质干细胞的可行性,可作为组织工程应用的潜在材料。

关键词: 海藻酸钠, 壳聚糖, 多聚赖氨酸, 间充质干细胞

Abstract: Objective Alginate-chitosan porous composite scaffold prepared and optimized was used to culture human umbilical cord-derived mesenchymal stem cells in order to provide the application of stem cells in tissue engineering and disease treatment. Methods We prepared porous composite scaffold, and which was optimized by grafting polylysine on its surface. Next, mesenchymal stem cells were isolated from umbilical cord, seeded and serially cultured in the scaffolds. Cell growth and proliferation were monitored, and cell proliferation curves were made using cytometry and CCK-8 kit techniques. Moreover, stem cell ageing was evaluated by senescence β-Galactosidase staining kit. Results This assay successfully prepared and optimized the alginate-chitosan composite scaffold. We found that the mesenchymal stem cells could easily attach to and cover the surface of the scaffold grafted with polylysine. Moreover, the stem cells grew in agglomerates rapidly. Conclusions This result proves that the alginate-chitosan scaffold provides a suitable environment for cell growth and has the feasibility for cell culture in vitro. This scaffold can be used as potential material for tissue engineering applications.

Key words: sodium alginate, chitosan, polylysine, mesenchymal stem cell