基于玻璃毛细管的骨骼肌细胞取向调控

Orientation Regulation of Skeletal Muscle Cells Based on Glass Capillary

目的基于玻璃毛细管侧壁对C2C12细胞的接触引导作用,提出一种低成本、高效率、便于批量使用的骨骼肌细胞取向调控新方法,并探究其应用价值。方法设计相关工艺合成甲基丙烯酸化明胶(GelMA),用于制备水凝胶薄膜和水凝胶纤维。以水凝胶薄膜为基底,以玻璃毛细管为调控结构对C2C12小鼠成肌细胞进行取向调控实验。研究了侧壁形状、基底和侧壁材料在细胞取向调控中的作用,探究此调控方法中真正决定细胞取向的因素。并基于原理对毛细管外径参数进行了优化实验。为说明调控所得细胞分化潜能,进行了分化与电刺激实验,将细胞分化形成肌管,施加外部电场并量化其响应效果。结果毛细管的外弧形侧壁对细胞调控作用明显,能得到垂直于毛细管轴向生长的大面积高密度细胞层。此调控方法中发挥主导因素的是侧壁形状,外径750μm毛细管具有最好的调控效果。调控所得细胞分化而成的肌管在外部电场下可产生有效的响应。结论玻璃毛细管的侧壁会引导细胞垂直毛细管轴向生长,可用于批量化骨骼肌细胞取向调控。此调控方法在组织工程和生物混合驱动等领域具有良好的应用前景。

Objective Based on the contact guiding function of the glass capillary side wall to C2 C12 cells,a low-cost,high-efficiency,and easy to batch use method for skeletal muscle cell orientation regulation was proposed and its application value was studied.Methods Methacrylate Gelatin(GelMA)was synthesized for the preparation of hydrogel films and hydrogel fibers.The C2 C12 cell orientation regulation experiment was carried out with the hydrogel film as the substrate and the glass capillary as the control structure.The roles of the side wall shape,the materials of the substrate and the control structure in the cell orientation regulation were studied to explore the key factors determining the cell orientation in this regulation method.Based on the regulation principle,an experiment was designed to optimize the capillary diameter parameters.Finally,the differentiation and electrical stimulation experiments were performed to illustrate the differentiation potential of the resulting cells.An external electric field was imposed on myotubes differentiated from the cells and their responses were quantified.Results The outer curved side walls of the capillaries had a good regulation effect on the cells,and produced a large-area high-density cell layer on which cells grew perpendicular to the axial direction of the capillaries.The results showed that the shape of the sidewall was the decisive factor in this method.It was also proved that the capillary with an outer diameter of 750μm had the best regulation effect.Those myotubes also produced an effective response in the electric field.Conclusion The side wall of the glass capillary could guide the cells to grow perpendicularly to the axial direction of the capillary which may be used to regulate the orientation of the skeletal muscle cells in batches.This regulation method has good application prospects in the fields of tissue engineering and bio-hybrid actuators.