Workman Victoria L, Tezera Liku B, Elkington Paul T, Jayasinghe Suwan N
BioPhysics Group, UCL Institute of Biomedical Engineering, UCL Centre for Stem Cells and Regenerative Medicine and Department of Mechanical Engineering, University College London, Torrington Place, London, WC1E 7JE, United Kingdom.
Clinical and Experimental Sciences, Institute for Life Sciences, Faculty of Medicine, University of Southampton, Southampton, SO16 6YD, United Kingdom.
Adv Funct Mater. 2014 May 14;24(18):2648-2657. doi: 10.1002/adfm.201303891.
A growing body of evidence suggests that studying cell biology in classical two-dimensional formats, such as cell culture plasticware, results in misleading, non-physiological findings. For example, some aspects of cancer biology cannot be observed in 2D, but require 3D culture methods to recapitulate observations in vivo. Therefore, we developed a microsphere-based model to permit 3D cell culture incorporating physiological extracellular matrix components. Bio-electrospraying was chosen as it is the most advanced method to produce microspheres, with THP-1 cells as a model cell line. Bio-electrospraying parameters, such as nozzle size, polymer flow rate, and voltage, were systematically optimized to allow stable production of size controlled microspheres containing extracellular matrix material and human cells. We investigated the effect of bio-electrospraying parameters, alginate type and cell concentration on cell viability using trypan blue and propidium iodide staining. Bio-electrospraying had no effect on cell viability nor the ability of cells to proliferate. Cell viability was similarly minimally affected by encapsulation in all types of alginate tested (MVM, MVG, chemical- and food-grade). Cell density of 5 × 10 cells ml within microspheres was the optimum for cell survival and proliferation. The stable generation of microspheres incorporating cells and extracellular matrix for use in a 3D cell culture will benefit study of many diverse diseases and permit investigation of cellular biology within a 3D matrix.
越来越多的证据表明，以传统二维形式（如细胞培养塑料制品）研究细胞生物学，会产生误导性的、非生理性的结果。例如，癌症生物学的某些方面在二维环境中无法观察到，而是需要三维培养方法来重现体内观察结果。因此，我们开发了一种基于微球的模型，以实现包含生理性细胞外基质成分的三维细胞培养。之所以选择生物电喷雾法，是因为它是生产微球最先进的方法，选用THP-1细胞作为模型细胞系。对生物电喷雾参数，如喷嘴尺寸、聚合物流速和电压进行了系统优化，以稳定生产尺寸可控的、含有细胞外基质材料和人类细胞的微球。我们使用台盼蓝和碘化丙啶染色研究了生物电喷雾参数、藻酸盐类型和细胞浓度对细胞活力的影响。生物电喷雾对细胞活力以及细胞增殖能力均无影响。在所有测试的藻酸盐类型（MVM、MVG、化学级和食品级）中，细胞包封对细胞活力的影响同样微乎其微。微球内5×10个细胞/毫升的细胞密度是细胞存活和增殖的最佳密度。稳定生成包含细胞和细胞外基质的微球用于三维细胞培养，将有利于多种疾病的研究，并有助于在三维基质中研究细胞生物学。
