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目的 研究主动脉平滑肌细胞 (ASMC)在仿真皮替代物的三维培养中是否能产生弹性纤维。 方法 取 1周龄SD大鼠胸主动脉 ,进行原代和传代培养 ,以α肌动蛋白免疫细胞化学染色鉴定平滑肌细胞。然后 ,将ASMC与胶原、甲壳素及糖胺多糖组合形成的凝胶混合 ,进行三维培养 ;培养 1周和 2周后 ,用Gomori醛复红染色和弹性蛋白免疫细胞化学染色 ,检测ASMC胶原凝胶。 结果 传代细胞经α肌动蛋白免疫细胞化学染色鉴定 ,98%以上为ASMC。培养 1周和 2周的ASMC胶原凝胶中 ,均显示有弹性纤维存在。 结论 ASMC在仿真皮替代物的三维培养中能够产生弹性蛋白 ,并形成弹性纤维。
Objective To investigate whether aortic smooth muscle cells (ASMC) produce elastic fibers in three-dimensional culture of dermal substitutes. Methods Thoracic aorta of 1-week-old SD rats were used for primary and subculture, and smooth muscle cells were identified by α-actin immunocytochemical staining. Then, ASMC was mixed with collagen gel, chitin and glycosaminoglycan to form three-dimensional culture. After cultured for 1 week and 2 weeks, Gomori aldehyde complex red staining and elastin immunocytochemistry staining were used to detect ASMC collagen coagulation glue. Results The passage cells were identified by α-actin immunocytochemistry. More than 98% of them were ASMC. ASMC collagen gels incubated for 1 and 2 weeks showed the presence of elastic fibers. Conclusion ASMC can produce elastin in three-dimensional culture of dermal substitute and form elastic fibers.