Goncharova Elena A, Lim Poay, Goncharov Dmitry A, Eszterhas Andrew, Panettieri Reynold A, Krymskaya Vera P
Pulmonary, Allergy & Critical Care Division, Department of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
Nat Protoc. 2006;1(6):2905-8. doi: 10.1038/nprot.2006.433.
Vascular and airway remodeling, which are characterized by airway smooth muscle (ASM) and pulmonary arterial vascular smooth muscle (VSM) proliferation, contribute to the pathology of asthma, pulmonary hypertension, restenosis and atherosclerosis. To evaluate the proliferation of VSM and ASM cells in response to mitogens, we perform a [3H]thymidine incorporation assay. The proliferation protocol takes approximately 48 h and includes stimulating cells synchronized in G0/G1 phase of the cell cycle with agonists, labeling cells with [3H]thymidine and examining levels of [3H]thymidine incorporation by scintillation counting. Although using radiolabeled [3H]thymidine incorporation is a limitation, the greatest benefit of the assay is providing reliable and statistically significant data.
血管和气道重塑以气道平滑肌（ASM）和肺动脉血管平滑肌（VSM）增殖为特征，是哮喘、肺动脉高压、再狭窄和动脉粥样硬化病理过程的一部分。为了评估VSM和ASM细胞对有丝分裂原的增殖反应，我们进行了[3H]胸苷掺入试验。增殖实验大约需要48小时，包括用激动剂刺激处于细胞周期G0/G1期的同步化细胞，用[3H]胸苷标记细胞，并通过闪烁计数检测[3H]胸苷掺入水平。尽管使用放射性标记的[3H]胸苷掺入存在局限性，但该试验的最大优点是能提供可靠且具有统计学意义的数据。
