Kato Koichi, Umezawa Kohei, Miyake Masato, Miyake Jun, Nagamune Teruyuki
National Institute of Advanced Industrial Science and Technology, Amagasaki, Japan.
Biotechniques. 2004 Sep;37(3):444-8, 450, 452. doi: 10.2144/04373RR02.
Cell-based microarrays are emerging as a tool for analyzing the functions of genes in cells. However, partly due to the difficulty of cell immobilization, the application of this method has been limited to adherent cells. We previously reported a method that rapidly and strongly attached living nonadherent cells to glass slides modified with a cell membrane anchoring reagent, designated a biocompatible anchor for membrane (BAM). Here we demonstrate that plasmid DNA deposited in a defined area on BAM-modified glass slides was transfected into nonadherent K562 cells immobilized on the DNA-deposited and BAM-modified slides. This method allowed the transfection of K562 cells not only with plasmid cDNA expression vectors but also with small interfering RNA (siRNA) at a defined location on the BAM-modified slides. We expect this methodology to greatly expand the scope of current cell microarray technology.
基于细胞的微阵列正逐渐成为一种分析细胞中基因功能的工具。然而,部分由于细胞固定的困难,该方法的应用仅限于贴壁细胞。我们之前报道了一种方法,可将活的非贴壁细胞快速且牢固地附着到用细胞膜锚定试剂修饰的载玻片上,该试剂称为膜生物相容性锚定物(BAM)。在此,我们证明沉积在BAM修饰载玻片特定区域的质粒DNA被转染到固定在DNA沉积且BAM修饰载玻片上的非贴壁K562细胞中。该方法不仅能使K562细胞在BAM修饰载玻片的特定位置用质粒cDNA表达载体转染,还能用小干扰RNA(siRNA)转染。我们期望这种方法能极大地扩展当前细胞微阵列技术的范围。