Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology, Central4 1-1-1 Higashi, Tsukuba, Ibaraki 305-8562, Japan.
Langmuir. 2013 May 28;29(21):6429-33. doi: 10.1021/la4012229. Epub 2013 May 16.
We report here a method for controlling cell adhesion, allowing simple yet accurate cell detachment from the substrate, which is required for the establishment of new cytometry-based cell processing and analyzing methods. A biocompatible anchor for membrane (BAM) was conjugated with bovine serum albumin (BSA) to produce a cell-anchoring agent (BAM-BSA). By coating polystyrene substrates with a mixture of BAM-BSA and BSA, controlled suppression of the substrate's adhesive properties was achieved. Hook-shaped nanoneedles were used to pick up cells from the substrate, while recording the cell-substrate adhesion force, using an atomic force microscope (AFM). Due to the lipid bilayer targeting property of BAM, the coated surface showed constant adhesion forces for various cell lines, and controlling the BAM-BSA/BSA ratio enabled tuning of the adhesion force, ranging from several tens of nano-Newtons down to several nano-Newtons. Optimized tuning of the adhesion force also enabled the detachment of cells from BAM-BSA/BSA-coated dishes, using a shear flow. Moreover, the method was shown to be noncell type specific and similar results were observed using four different cell types, including nonadherent cells. The attenuation of cell adhesion was also used to enable the collection of single cells by capillary aspiration. Thus, this versatile and relatively simple method can be used to control the adhesion of various cell types to substrates.
我们在此报告一种控制细胞黏附的方法，该方法可简单而精确地实现细胞从基底上的脱离，这是建立新的基于细胞检测的细胞处理和分析方法所必需的。将生物相容性的膜锚（BAM）与牛血清白蛋白（BSA）偶联以产生细胞锚定剂（BAM-BSA）。通过将 BAM-BSA 和 BSA 的混合物涂覆在聚苯乙烯基底上，实现了对基底粘合性能的有效抑制。使用原子力显微镜（AFM），通过钩状纳米针从基底上拾取细胞，同时记录细胞-基底的黏附力。由于 BAM 具有靶向脂质双层的特性，因此涂覆的表面对各种细胞系均表现出恒定的黏附力，并且通过控制 BAM-BSA/BSA 的比例，可以调节黏附力，范围从数十毫牛顿到数毫牛顿。通过优化黏附力的调节，还可以使用切向流从 BAM-BSA/BSA 涂覆的培养皿中分离细胞。此外，该方法显示出非细胞类型特异性，并且在四种不同的细胞类型（包括非黏附细胞）中观察到了相似的结果。细胞黏附的减弱还可用于通过毛细吸引收集单细胞。因此，这种多功能且相对简单的方法可用于控制各种细胞类型与基底的黏附。
