生物学杂志 ›› 2020, Vol. 37 ›› Issue (5): 30-.doi: 10.3969/j.issn.2095-1736.2020.05.030

• 研究报告 • 上一篇    下一篇

纳米细菌纤维素复合物支架用于MDCK细胞三维培养的研究

  

  1. 1. 武汉大学 口腔医院, 武汉 430079; 2. 武汉大学 药学院, 武汉 430071;3. 中南民族大学 生命科学学院, 武汉 430074
  • 出版日期:2020-10-18 发布日期:2020-10-14
  • 通讯作者: 曾晓波,副教授,研究方向为微生物制药,E-mail: zengxb@ whu.edu.cn
  • 作者简介:关业茂,主管药师,研究方向为药物靶向治疗,E-mail: 936977161@qq.com
  • 基金资助:
    中南民族大学中央高校基本科研基金(基金编号CZT18001)

Study on nanobacterial cellulose complex scaffold forthree-dimensional culture of MDCK cells

  1. 1. Stomatological Hospital of Wuhan University, Wuhan 430079;2. School of Pharmaceutical Sciences, Wuhan University, Wuhan 430071;3. School of Life Sciences, South-Central University for Nationalities, Wuhan 430074, China
  • Online:2020-10-18 Published:2020-10-14

摘要: 三维细胞培养广泛应用于生物医学研究的各个领域,三维培养支架材料的研究是其中重要方面。细菌来源的纤维素经酸水解得到纳米纤维素,电镜观察其长度为600~800 nm,直径为50~150 nm。将其分别与海藻酸钠和Matrigel复合制成三维培养支架,测试其对MDCK细胞三维培养的效果。发现单独使用海藻酸钠作为凝胶支架材料,MDCK细胞在其中生长状况较差。纳米细菌纤维素与海藻酸钠复合制成的凝胶支架,可以提高MDCK细胞的生长,形成较多细胞团。对复合物支架上形成的细胞团进行细胞核与细胞膜荧光染色后观察,细胞团呈现三维生长所特有的囊腔结构,说明纳米细菌纤维素与海藻酸钠复合物支架可以用于MDCK细胞三维培养。在复合物中添加0.5%的Matrigel制成的凝胶支架,可以明显促进MDCK细胞的三维生长。这种在复合物中添加少量Matrigel以补充细胞三维生长必需的胞外基质的方式,在应用开发上具有一定前景。研究为探索纳米细菌纤维素作为细胞三维培养支架材料提供了前期基础。

关键词: 三维培养, 支架材料, 纳米细菌纤维素, 海藻酸钠

Abstract: Three-dimensional cell culture is widely used in various fields of biomedical research. The research of three-dimensional culture scaffold materials is an important aspect. Nanocellulose with a length of 600-800 nm and a diameter of about 50-150 nm was prepared from bacterial cellulose by acidic hydrolyzing. The nanobacterial cellulose was combined with sodium alginate and Matrigel to prepare a three-dimensional culture scaffold to test its effect on MDCK cells′ three-dimensional growth. It was found that MDCK cells grew poorly in the gel scaffold only containing sodium alginate. A scaffold combining nanobacterial cellulose and sodium alginate could increase the growth of MDCK cells and form more cell clusters. Fluorescent staining nucleus and cell membrane of the cell cluster formed in the composite scaffold showed the MDCK cell mass exhibited a cystic structure unique to three-dimensional growth. This indicated that the nanobacterial cellulose and sodium alginate complex scaffold could be used for three-dimensional culture of MDCK cells. Adding 0.5% Matrigel to the complex significantly promoted the three-dimensional growth of MDCK cells. This method of adding a small amount of Matrigel to the complex to supplement the extracellular matrix necessary for three-dimensional growth of cells had certain prospects in application development. The research would provide a preliminary basis for exploring nanobacterial cellulose as a scaffold material for three-dimensional cell culture.

Key words: three-dimensional culture, scaffold material, nanobacterial cellulose, sodium alginate

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