Centre for Cell Engineering, Institute of Molecular, Cellular and Systems Biology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UK.
J Tissue Eng Regen Med. 2012 Dec;6 Suppl 3:s4-14. doi: 10.1002/term.510. Epub 2011 Dec 13.
In tissue engineering, chemical and topographical cues are normally developed using static cell cultures but then applied directly to tissue cultures in three dimensions (3D) and under perfusion. As human cells are very sensitive to changes in the culture environment, it is essential to evaluate the performance of any such cues in a perfused environment before they are applied to tissue engineering. Thus, the aim of this research was to bridge the gap between static and perfusion cultures by addressing the effect of perfusion on cell cultures within 3D scaffolds. For this we developed a scaled-down bioreactor system, which allows evaluation of the effectiveness of various chemical and topographical cues incorporated into our previously developed tubular ε-polycaprolactone scaffold under perfused conditions. Investigation of two exemplary cell types (fibroblasts and cortical astrocytes) using the miniaturized bioreactor indicated that: (a) quick and firm cell adhesion in the 3D scaffold was critical for cell survival in perfusion culture compared with static culture; thus, cell-seeding procedures for static cultures might not be applicable, therefore it was necessary to re-evaluate cell attachment on different surfaces under perfused conditions before a 3D scaffold was applied for tissue cultures; (b) continuous medium perfusion adversely influenced cell spread and survival, which could be balanced by intermittent perfusion; (c) micro-grooves still maintained their influences on cell alignment under perfused conditions, while medium perfusion demonstrated additional influence on fibroblast alignment but not on astrocyte alignment on grooved substrates. This research demonstrated that the mini-bioreactor system is crucial for the development of functional scaffolds with suitable chemical and topographical cues by bridging the gap between static culture and perfusion culture.
在组织工程中，通常使用静态细胞培养来开发化学和拓扑线索，然后直接将其应用于三维（3D）和灌注下的组织培养中。由于人类细胞对培养环境的变化非常敏感，因此在将这些线索应用于组织工程之前，必须在灌注环境中评估任何此类线索的性能。因此，本研究的目的是通过解决灌注对 3D 支架内细胞培养的影响来弥合静态和灌注培养之间的差距。为此，我们开发了一个缩小规模的生物反应器系统，该系统允许在灌注条件下评估我们之前开发的管状 ε-聚己内酯支架中纳入的各种化学和拓扑线索的有效性。使用微型生物反应器研究两种典型的细胞类型（成纤维细胞和皮质星形胶质细胞）表明：（a）与静态培养相比，灌注培养中 3D 支架中快速而牢固的细胞黏附对于细胞存活至关重要；因此，静态培养中的细胞接种程序可能不适用于灌注培养，因此在将 3D 支架应用于组织培养之前，有必要在灌注条件下重新评估不同表面上的细胞黏附；（b）连续的培养基灌注会对细胞扩散和存活产生不利影响，可以通过间歇灌注来平衡；（c）微槽在灌注条件下仍保持对细胞排列的影响，而培养基灌注对成纤维细胞的排列有额外的影响，但对槽状基底上的星形胶质细胞的排列没有影响。这项研究表明，微型生物反应器系统对于通过弥合静态培养和灌注培养之间的差距来开发具有合适化学和拓扑线索的功能性支架至关重要。
