Kuschel Cornelia, Steuer Heiko, Maurer Andreas N, Kanzok Britta, Stoop Reinout, Angres Brigitte
NMI at the University of Tübingen, Reutlingen, Germany.
Biotechniques. 2006 Apr;40(4):523-31. doi: 10.2144/000112134.
We have developed a microarray-based system for cell adhesion profiling of large panels of cell-adhesive proteins to increase the throughput of in vitro cell adhesion assays, which are currently primarily performed in multiwell plates. Miniaturizing cell adhesion assays to an array format required the development of protocols for the reproducible microspotting of extracellular matrix (ECM) protein solutions and for the handling of cell suspensions during the assay. We generated ECM protein microarrays with high reproducibility in microspot protein content using nitrocellulose-coated glass microslides, combined with piezoelectric microspotting of protein solutions. Protocols were developed that allowed us to use 5000 cells or fewer on an array of 4 x 4 mm consisting of 64 microspots. Using this microarray system, we identified differences of adhesive properties of three cell lines to 14 different ECM proteins. Furthermore, the sensitivity and accuracy of the assays were increased using microarrays with ranges of ECM protein amounts. This microarray system will be particularly useful for extensive comparative cell adhesion profiling studies when only low amounts of adhesive substrate and cells, such as stem cells or cells from biopsies, are available.
我们开发了一种基于微阵列的系统，用于对大量细胞粘附蛋白进行细胞粘附分析，以提高体外细胞粘附试验的通量，目前该试验主要在多孔板中进行。将细胞粘附试验小型化为阵列形式需要开发细胞外基质（ECM）蛋白溶液可重复微点样以及试验过程中处理细胞悬液的方案。我们使用硝酸纤维素包被的玻璃微载玻片，结合蛋白溶液的压电微点样，生成了微点蛋白含量具有高重现性的ECM蛋白微阵列。开发的方案使我们能够在由64个微点组成的4×4 mm阵列上使用5000个或更少的细胞。使用该微阵列系统，我们鉴定了三种细胞系对14种不同ECM蛋白的粘附特性差异。此外，使用具有不同ECM蛋白量范围的微阵列提高了试验的灵敏度和准确性。当只有少量粘附底物和细胞（如干细胞或活检组织中的细胞）可用时，这种微阵列系统对于广泛的比较细胞粘附分析研究将特别有用。
