三维培养状态下肌腱细胞骨架对细胞生物学行为的影响

目的 探讨肌腱细胞在三维支架上培养方式下细胞骨架的结构分布对细胞生物学行为的影响。方法将肌腱细胞与梯度降解肌腱支架材料(GDBM)体外复合三维培养，进行细胞骨架形态学观察、细胞增殖、细胞周期、细胞DNA倍体水平及肌腱细胞凋亡检测。结果GDBM组细胞第2天进入对数增长期，倍增时间为3．25d，而单纯培养对照组倍增时间为3．75d。与支架材料复合培养的肌腱细胞DNA指数为0．96，增殖指数比对照组高2．1％，其细凋亡率明显低于二维培养。提示肌腱细胞在三维支架上生长速度快，增殖能力强。结论三维培养状态下肌腱细胞在梯度降解肌腱支架材料上生长良好，梯度降解肌腱支架材料具有良好的生物相容性，可作为肌腱细胞的有效载体应用于组织工程化肌腱的构建。

Experimental studies of influence of cytoskeleton on tenocyte biological behavior under three-dimensional culture

Objective To study the influence of the cytoskeleton on biological behavior of cultured tenocytes under three dimensional scaffold.Methods Tenocyes were separated from Roman chicken's flextor digitorum profoundus,and cultured with gradient degradation biomaterials (GDBM) in vitro.The morphological cytoskeleton characters,cell proliferation,mitotic cycle,DNA index and apoptosis were detected.Results The tenocytes attached to GDBM and grew along fibers in spindle like manner.Cells in GDBM group reached logarithm increasing period at the second day.Its doubling time was 3.25 day,while that of the cells in control group was 3.75 day.The DNA index of cells in GDBM group was 0.96,proliferation index 2.1% higher than in the control group,apoptosis rates lower than plate culture ( P < 0.05), indicating that the tenocytes cultured with GDBM grew and proliferated faster.Conclusion GDBM show good biocompatibility with tenocytes and they were promising extracellular matrix scaffold for cell transplantation in tendon tissue engineering.