Schwann细胞与硫酸肝素-胶原蛋白支架材料的复合

COMPOUND OF SCHWANN CELLS IN HEPARIN SULFATE-COLLAGEN SCAFFOLD

本研究目的是观测硫酸肝素-胶原蛋白支架材料对Schwann细胞的相容性,采用: (1)冷冻干燥法制备硫酸肝素-胶原蛋白支架材料,培养、纯化并鉴定Schwann细胞后,将Hoechst荧光标记的Schwann细胞复合于硫酸肝素-胶原支架材料中,荧光显微镜、扫描电镜及光镜下观察其在材料内部的空间排列形式; (2)以台盼蓝染色、MTT法检测Schwann细胞与硫酸肝素-胶原蛋白共培养时的生物活性。结果显示:作者制备的支架材料具有纵行的、平行排列的微管结构,Schwann细胞在支架材料微管内成线性平行排列,类似于神经基底膜与Schwann细胞形成的Büngner带;台盼蓝染色Schwann细胞死亡率仅6. 47%,MTT法证明Schwann细胞与硫酸肝素-胶原蛋白材料共培养时保有高度的生物活性。本研究结果表明:以硫酸肝素复合胶原蛋白结合特定条件下冷冻干燥技术可以制备在组分和内部空间结构上高度仿生神经的新型神经组织工程支架材料,与Schwann细胞有良好的细胞相容性。

To observe the compatibility of heparin sulfate-collagen scaffold with Schwann cells. Heparin sulfate-collagen scaffold was produced by freeze-drying. After the Schwann cells were cultured, purified and indentified, they were labeled with Hoechst fluorescence and compounded on heparin sulfate-collagen scaffold. The arrangement of the cells was observed through fluorescence microscope, scanning electron microscope and light microscope. Bioactivity of Schwann cells when co-culturing together with heparin sulfate-collagen scaffold was inspected by trypan blue staining and MTT method. Heparin sulfate-collagen scaffold has longitudinal and parallel microtubule structure. Schwann cells grew in linear paralleling pattern, much similar with Büngner band formed by Schwann cell and nerve basement membrane. Trypan blue staining showed that the Schwann cell death rate was only 6.47%. MTT detection showed that Schwann cells preserved bioactivity while co-culturing with heparin sulfate-collagen scaffold. Heparin sulfate-collagen scaffold, produced under special freeze-drying condition, is a new kind of nervous tissue engineering scaffold, which is highly mimic natural nerve in space structure and component and has nice cellular compatibility with Schwann cells.