The use of non-bronchoscopic brushings to study the paediatric airway
The use of non-bronchoscopic brushings to study the paediatric airway
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DOI：
10.1186/1465-9921-6-53
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发表时间：
2005-06-08
影响因子：
5.8
通讯作者：
Stick, S
中科院分区：
文献类型：
--
作者：
Lane, C;Burgess, S;Stick, S
背景资料：使用细胞学刷从成人获得呼吸道细胞用于临床和研究目的是公认的。然而，尚未评估非支气管镜刷拭用于研究儿科气道的安全性和实用性。本研究的目的是评估使用非支气管镜刷拭对儿童上皮细胞进行采样的实用性，以使用广泛的分子和细胞技术研究健康和疾病中的上皮功能。在一个非选择的健康队列中研究了非支气管镜刷牙，轻度哮喘儿童因与呼吸系统疾病无关的手术而就诊，在珀斯的主要儿童医院。对手术的安全性和副作用进行了评估。测量所有样品的细胞数量、表型和活力。这些细胞用于长期细胞培养，免疫组化，蛋白质印迹，定量PCR和基因阵列的潜力进行了检查。结果：非支气管镜刷牙耐受性良好，在所有儿童。手术后唯一显著的副作用是咳嗽：护理人员报告20%的患者咳嗽;父母报告40%的患者咳嗽。取样的细胞数量和质量足以在93%的样品中进行细胞培养。同样，从细胞中提取的蛋白质和RNA是合适的基因和蛋白质表达的调查，使用微阵列和实时PCR.Conclusion：非支气管镜刷在儿童是安全的，易于执行，是不相关的任何并发症。使用该技术，可以回收足够数量的上皮细胞以允许细胞培养、蛋白质印迹、真实的时间PCR和微阵列分析。本研究的目的是证明非支气管镜气道刷拭在获得和研究上皮细胞方面的实用性，并鼓励其他人，以便我们能够加速了解上皮细胞在儿童呼吸道疾病中的作用。
Background: The use of cytology brushes for the purpose of obtaining respiratory cells from adults for clinical and research purposes is well established. However, the safety and utility of non-bronchoscopic brushings to study the paediatric airway has not been assessed. The purpose of this study was to assess the practicality of using non-bronchoscopic brushing to sample epithelial cells from children for investigation of epithelial function in health and disease using a wide range of molecular and cellular techniques.Methods: Non-bronchoscopic brushing was investigated in a non-selected cohort of healthy, and mildly asthmatic children presenting for surgery unrelated to respiratory conditions, at the major children's hospital in Perth. Safety and side-effects of the procedure were assessed. Cell number, phenotype and viability were measured for all samples. The potential of these cells for use in long-term cell culture, immunohistochemistry, western blotting, quantitative PCR and gene arraying was examined.Results: Non-bronchoscopic brushing was well tolerated in all children. The only significant side effect following the procedure was cough: nursing staff reported cough in 20% of patients; parents reported cough in 40% of patients. Cells sampled were of sufficient quantity and quality to allow cell culture in 93% of samples. Similarly, protein and RNA extracted from the cells was suitable for investigation of both gene and protein expression using micro-array and real-time PCR.Conclusion: Non-bronchoscopic brushing in children is safe and easy to perform, and is not associated with any complications. Using this technique, adequate numbers of epithelial cells can be retrieved to allow cell culture, western blotting, real time PCR, and microarray analysis. The purpose of this study is to demonstrate the utility of non-bronchoscopic airway brushing to obtain and study epithelial cells and to encourage others so that we can accelerate our knowledge regarding the role of the epithelium in childhood respiratory disease.