中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (21): 3827-3831.doi: 10.3969/j.issn.1673-8225.2010.21.007

• 细胞外基质材料 extracellular matrix materials • 上一篇    下一篇

碱性成纤维细胞生长因子对许旺细胞在小肠黏膜下层支架上黏附及增殖的影响

张开伟1，肖  睿2，段  宏1，项  舟1，3，李秀群3   

  1. 1四川大学华西医院骨科，四川省成都市   610041；2宜宾市第一人民医院骨科，四川省宜宾市，644000；3四川大学华西医院生物治疗国家重点实验室干细胞与组织工程研究室，四川省成都市  610041
  • 出版日期:2010-05-21 发布日期:2010-05-21
  • 通讯作者: 项 舟，教授，主任医师，博士生导师，四川大学华西医院骨科，四川省成都市 610041 xiangzhou15@hotmail.com
  • 作者简介:张开伟☆，男，1973年生，贵州省贵阳市人，汉族，华西临床医学院在读博士，副主任医师，主要从事骨与关节损伤方面的研究。 zkw1973@yahoo.com.cn
  • 基金资助:

    四川省宜宾市重点科技计划项目（200803010）。

Effect of basic fibroblast growth factor on adhesion and proliferation of Schwann cells on small intestinal submucosa scaffold

Zhang Kai-wei1, Xiao Rui2, Duan Hong1, Xiang Zhou 1,3, Li Xiu-qun3   

  1. 1 Department of Orthopaedics, West China Hospital, Sichuan University, Chengdu   610041, Sichuan Province, China; 2 Department of Orthopaedics, the First People’s Hospital of Yibin, Yibin  644000, Sichuan Province, China; 3 Division of Stem Cells and Tissue Engineering, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu   610041, Sichuan Province, China
  • Online:2010-05-21 Published:2010-05-21
  • Contact: Xiang Zhou, Doctor, Professor, Chief physician, Doctoral supervisor, Department of Orthopaedics, West China Hospital, Sichuan University, Chengdu 610041, Sichuan Province, China; Division of Stem Cells and Tissue Engineering, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, Sichuan Province, China xiangzhou15@hotmail.com
  • About author:Zhang Kai-wei☆, Studying for doctorate, Associate chief physician, Department of Orthopaedics, West China Hospital, Sichuan University, Chengdu 610041, Sichuan Province, China zkw1973@yahoo.com.cn
  • Supported by:

    the Key Science and Technology Development Program of Sichuan Province, No. 200803010*

PDF

333

被引次数

4

摘要：

背景：许旺细胞复合小肠黏膜下层是构建人工神经的可行方法，碱性成纤维细胞生长因子有促进许旺细胞增殖的作用。
目的：验证碱性成纤维细胞生长因子对许旺细胞在小肠黏膜下层支架材料表面的细胞增殖及黏附状态的影响。
方法：将体外分离培养的2代SD乳鼠许旺细胞接种于小肠黏膜下层支架材料上并加入50 μg/L的碱性成纤维细胞生长因子复合培养为实验组，以单纯的许旺细胞复合小肠黏膜下层作为对照组，分别用MTT法测定细胞增殖能力，细胞黏附率检测细胞在支架上的黏附情况，用流式细胞仪测定细胞分裂周期，并用苏木精-伊红染色及描电镜观察细胞形态及与材料的贴附情况。
结果与结论：MTT显示实验组的细胞增殖吸光度值明显高于对照组(P < 0.05)，实验组的细胞黏附率为(69.47±3.17)%，对照组为(44.58±1.76)%(P < 0. 05)，细胞周期显示实验组比对照组的G2/M+S期细胞百分含量高(P < 0.05)，培养7 d后的苏木精-伊红染色及扫描电镜显示实验组比对照组材料上聚集的细胞数量多，细胞形态伸展更明显，细胞贴附力更好。因此碱性成纤维细胞生长因子可明显地改善许旺细胞在小肠黏膜下层上的增殖及黏附能力，能促进许旺细胞与小肠黏膜下层支架的复合而构建人工神经导管。

关键词: 碱性成纤维细胞生长因子, 许旺细胞, 小肠黏膜下层, 增殖, 黏附

Abstract:

BACKGROUND: Schwann cells combined with small intestinal submucosa (SIS) is a feasible way to construct artificial nerve. Basic fibroblast growth factor (bFGF) can promote proliferation of Schwann cells.
OBJECTIVE: To explore the effects of bFGF on adhesion and proliferation of Schwann cells on scaffold of SIS.
METHODS: The second passage of Schwann cells isolated from neonatal SD rats were seeded onto the surface of SIS scaffold and incubated with 50 μg/L bFGF, which served as experimental group. The cultures of SC and SIS without bFGF were used as control. The cell proliferation was measured with MTT assay, and the adhesion of Schwann cells was determined by the rate of cell adhesion; the cell cycle of SC was examined by flow cytometry. The morphology of cells and cell adhesion on SIS were observed by contrast phase microscope, hematoxylin-eosin (HE) staining and scanning electron microscopy.
RESULTS AND CONCLUSION: Absorbance value of SC in experimental group was higher than the control group (P < 0.05). The cell adhesion rate was (69.47±3.17)% and (44.58±1.76)% respectively for experimental and control groups (P < 0.05). G2/M+S percentage of SCs was significantly greater in experimental group than the control group (P < 0.05). HE staining and microscopy showed that the number of attached cells was more and cell adhesion was better in experimental group compared with the control group at 7 days after culture. bFGF is effective in improving the adhesion and proliferation of SCs on SIS scaffold, and also promoting construction of tissue-engineered nerve conduit based on the combination with SC and SIS.

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