Meyer Rachel, Wong Wen Yu, Guzman Roberto, Burd Randy, Limesand Kirsten
Department of Nutritional Sciences, University of Arizona.
Cancer Biology Graduate Interdisciplinary Program, University of Arizona.
J Vis Exp. 2019 May 17(147). doi: 10.3791/59484.
Hyposalivation and xerostomia create chronic oral complications that decrease the quality of life in head and neck cancer patients who are treated with radiotherapy. Experimental approaches to understanding mechanisms of salivary gland dysfunction and restoration have focused on in vivo models, which are handicapped by an inability to systematically screen therapeutic candidates and efficiencies in transfection capability to manipulate specific genes. The purpose of this salivary gland organotypic culture protocol is to evaluate maximal time of culture viability and characterize cellular changes following ex vivo radiation treatment. We utilized immunofluorescent staining and confocal microscopy to determine when specific cell populations and markers are present during a 30-day culture period. In addition, cellular markers previously reported in in vivo radiation models are evaluated in cultures that are irradiated ex vivo. Moving forward, this method is an attractive platform for rapid ex vivo assessment of murine and human salivary gland tissue responses to therapeutic agents that improve salivary function.
唾液分泌减少和口干会引发慢性口腔并发症,降低接受放射治疗的头颈癌患者的生活质量。了解唾液腺功能障碍和恢复机制的实验方法主要集中在体内模型上,这些模型存在无法系统筛选治疗候选药物以及转染能力有限无法操纵特定基因的缺陷。本唾液腺器官型培养方案的目的是评估培养物的最大存活时间,并表征离体放射治疗后的细胞变化。我们利用免疫荧光染色和共聚焦显微镜来确定在30天的培养期内特定细胞群体和标志物何时出现。此外,在离体照射的培养物中评估先前在体内放射模型中报道的细胞标志物。展望未来,该方法是一个有吸引力的平台,可用于快速离体评估小鼠和人类唾液腺组织对改善唾液功能的治疗药物的反应。