Blau A, Weinl C, Mack J, Kienle S, Jung G, Ziegler C
Division of Biology 156-29, California Institute of Technology, Pasadena, CA 91125, USA.
J Neurosci Methods. 2001 Nov 15;112(1):65-73. doi: 10.1016/s0165-0270(01)00458-7.
New strategies for spatially controllable cell adhesion have been developed for brain cells from embryonic chicken. They are based on electrochemically active phenol and pyrrole derivatives, and can be used for the selective coverage of electroconductive substrates. Besides mimicking standard laminin-related adhesion promoting mechanisms by means of an electroactive monomer-linked 18-peptide segment from laminin (SRARKQAASIKVAVSADR), electrochemically generated thin (6-30 nm) polymer films of 3-hydroxybenzyl-hydrazine (3HBH) and 2-(3-hydroxyphenyl)-ethanol (2(3HP)E) with and without mechanically entrapped or covalently linked D-lysine have proved to promote cell adhesion in serum-free medium on indium-doped tin oxide (ITO) substrates during the first 6 culturing days in vitro. The effectiveness of the peptide was strongly density-dependent. Unexpectedly, laminin itself or a combination of laminin and poly-D-lysine (PDL) did not promote cell adhesion and neuron differentiation in serum-free cultures on ITO. However, they worked perfectly well on regular polystyrene substrates in serum-free medium or on ITO when medium with serum was used. This finding might suggest that the adhesion efficiency of laminin does not depend only on the kind of medium supplement but also on the type of substrate. In contrast, the adhesion-promoting properties of "artificial" polymeric films seemed to be based on a more direct cell-film interaction, with the film masking the substrate properties.
针对鸡胚胎脑细胞，已经开发出了空间可控细胞黏附的新策略。这些策略基于具有电化学活性的苯酚和吡咯衍生物，可用于选择性覆盖导电基底。除了通过层粘连蛋白的电活性单体连接的18肽段（SRARKQAASIKVAVSADR）模拟标准的层粘连蛋白相关黏附促进机制外，已证明，在体外培养的前6天，含和不含机械包裹或共价连接的D-赖氨酸的3-羟基苄基肼（3HBH）和2-（3-羟基苯基）乙醇（2(3HP)E）的电化学生成的薄（6-30纳米）聚合物膜，能促进无血清培养基中铟掺杂氧化锡（ITO）基底上的细胞黏附。该肽的有效性强烈依赖于密度。出乎意料的是，在ITO上的无血清培养中，层粘连蛋白本身或层粘连蛋白与聚-D-赖氨酸（PDL）的组合并不能促进细胞黏附和神经元分化。然而，在无血清培养基中的常规聚苯乙烯基底上或使用含血清培养基时在ITO上，它们的效果非常好。这一发现可能表明，层粘连蛋白的黏附效率不仅取决于培养基补充物的种类，还取决于基底的类型。相比之下，“人工”聚合物膜的黏附促进特性似乎基于更直接的细胞-膜相互作用，膜掩盖了基底的特性。
