Department of Pulmonary Medicine, Beijing Tian Tan Hospital, Capital Medical University, Beijing, China.
Respiration. 2013;85(6):500-4. doi: 10.1159/000346648. Epub 2013 Mar 27.
Airway granulation tissue and scar formation pose a challenge because of the high incidence of recurrence after treatment. As an emerging treatment modality, topical application of mitomycin C has potential value in delaying the recurrence of airway obstruction. Several animal and clinical studies have already proven its feasibility and efficacy. However, the ideal dosage has still not been determined.
To establish a novel method for culturing primary fibroblasts isolated from human airway granulation tissue, and to investigate the dose-effect of mitomycin C on the fibroblast proliferation in vitro, so as to provide an experimental reference for clinical practitioners.
Granulation tissues were collected during the routine bronchoscopy at our department. The primary fibroblasts were obtained by culturing the explanted tissues. The cells were treated with different concentrations of mitomycin C (0.1, 0.2, 0.4, 0.8 and 1.6 mg/ml) for 5 min followed by additional 48-hour culture before an MTT assay was performed to measure cell viability.
MTT assay showed that mitomycin C reduced cell viability at all tested concentrations. The inhibitory ratios were 10.26, 26.77, 32.88, 64.91 and 80.45% for cells treated with mitomycin C at 0.1, 0.2, 0.4, 0.8 and 1.6 mg/ml, respectively.
Explant culture is a reliable method for culturing primary fibroblasts from human airway granulation tissue, and mitomycin C can inhibit proliferation of the fibroblasts in vitro.
气道肉芽组织和瘢痕形成是一个挑战,因为治疗后复发率很高。作为一种新兴的治疗方式,丝裂霉素 C 的局部应用在延迟气道阻塞复发方面具有潜在价值。一些动物和临床研究已经证明了其可行性和疗效。然而,理想的剂量尚未确定。
建立一种培养人气道肉芽组织原代成纤维细胞的新方法,并研究丝裂霉素 C 对成纤维细胞体外增殖的剂量效应,为临床医生提供实验参考。
在我院常规支气管镜检查中收集肉芽组织。通过培养离体组织获得原代成纤维细胞。用不同浓度的丝裂霉素 C(0.1、0.2、0.4、0.8 和 1.6 mg/ml)处理细胞 5 分钟,然后再培养 48 小时,然后进行 MTT 测定以测量细胞活力。
MTT 测定表明,丝裂霉素 C 在所有测试浓度下均降低了细胞活力。用丝裂霉素 C 处理浓度为 0.1、0.2、0.4、0.8 和 1.6 mg/ml 的细胞的抑制率分别为 10.26%、26.77%、32.88%、64.91%和 80.45%。
组织块培养是培养人气道肉芽组织原代成纤维细胞的可靠方法,丝裂霉素 C 可以抑制成纤维细胞的体外增殖。