North East England Stem Cell Institute (NESCI), School of Biological and Biomedical Science, University of Durham, South Road, Durham, DH1 3LE, UK.
Cytotechnology. 2008 Feb;56(2):71-9. doi: 10.1007/s10616-007-9119-7. Epub 2008 Jan 25.
Many factors contribute to the creation and maintenance of a realistic environment for cell growth in vitro, e.g. the consistency of the growth medium, the addition of supplements, and the surface on which the cells grow. The nature of the surface on which cells are cultured plays an important role in their ability to attach, proliferate, migrate and function. Components of the extracellular matrix (ECM) are often used to coat glass or plastic surfaces to enhance cell attachment in vitro. Fragments of ECM molecules can be immobilised on surfaces in order to mimic the effects seen by whole molecules. In this study we evaluate the application of a novel technology for the immobilisation of functional domains of known ECM proteins in a controlled manner on a surface. By examining the adherence of cultured PC12 cells to alternative growth surfaces, we show that surfaces coated with motifs from collagen I, collagen IV, fibronectin and laminin can mimic surfaces coated with the corresponding whole molecules. Furthermore, we show that the adherence of cells can be controlled by modifying the hydropathic properties of the surface to either enhance or inhibit cell attachment. Collectively, these data demonstrate the application of a new technology to enable optimisation of cell growth in the tissue culture laboratory.
许多因素促成了细胞在体外生长的真实环境的创造和维持,例如生长培养基的一致性、添加的补充剂以及细胞生长的表面。细胞培养表面的性质对其附着、增殖、迁移和功能能力起着重要作用。细胞外基质 (ECM) 的成分通常用于涂覆玻璃或塑料表面,以增强体外细胞附着。可以将 ECM 分子的片段固定在表面上,以模拟整个分子所产生的效果。在这项研究中,我们评估了一种新技术在可控条件下将已知 ECM 蛋白的功能域固定在表面上的应用。通过检查培养的 PC12 细胞对替代生长表面的粘附,我们表明,涂有来自胶原蛋白 I、胶原蛋白 IV、纤连蛋白和层粘连蛋白的基序的表面可以模拟涂有相应整个分子的表面。此外,我们表明可以通过修饰表面的疏水性来控制细胞的粘附,以增强或抑制细胞附着。总的来说,这些数据证明了新技术的应用能够优化组织培养实验室中的细胞生长。