Laboratoire de Chimie Analytique et Bromatologie, EA 3842, Homéostasie Cellulaire et Pathologies, Université de Limoges, Faculté de Pharmacie, 2 rue du Docteur Marcland, 87025 Limoges Cedex, France.
J Chromatogr A. 2009 Dec 25;1216(52):9125-33. doi: 10.1016/j.chroma.2009.08.042. Epub 2009 Aug 21.
Classically described as a macroscale size-density based method, Sedimentation field flow fractionation (SdFFF) has been successfully used for cell sorting. The goal of this study was to develop a new SdFFF device for downscale applications, in particular for oncology research to rapidly monitor chemical biological event induction in a cell line. The development of a downscale SdFFF device required reduction of the separation channel volume. Taking advantage of a newly laboratory designed apparatus, channel volume was successfully decreased by reducing both length and breadth. To validate the apparatus and method, we used the well-known model of diosgenin dose-dependent induction of apoptosis or megakaryocytic differentiation in HEL cells. After a minute scale acquisition of a reference profile, the downscale device was able to perform fast, early, significant and reproducible monitoring of apoptosis and differentiation, two important biological mechanisms in the field of cancer research.
经典的沉降场流分离(SdFFF)是一种基于宏观尺寸和密度的方法,已成功用于细胞分选。本研究的目的是开发一种用于缩小规模应用的新型 SdFFF 设备,特别是用于肿瘤学研究,以快速监测细胞系中化学-生物事件的诱导。缩小 SdFFF 设备的规模需要减小分离通道的体积。利用新设计的实验室设备,通过减小长度和宽度,成功地减小了通道体积。为了验证设备和方法,我们使用了众所周知的薯蓣皂苷元剂量依赖性诱导 HEL 细胞凋亡或巨核细胞分化的模型。在获取一分钟尺度的参考图谱后,该小型设备能够快速、早期、显著且可重复地监测凋亡和分化,这是癌症研究领域的两个重要生物学机制。