Chin Kyuongsik, Khattak Sarwat F, Bhatia Surita R, Roberts Susan C
Department of Chemical Engineering, University of Massachusetts, 686 North Pleasant Street, Amherst, MA 01003, USA.
Biotechnol Prog. 2008 Mar-Apr;24(2):358-66. doi: 10.1021/bp070160f. Epub 2008 Feb 23.
Cell encapsulation provides cells a three-dimensional structure to mimic physiological conditions and improve cell signaling, proliferation, and tissue organization as compared to monolayer culture. Encapsulation devices often encounter poor mass transport, especially for oxygen, where critical dissolved levels must be met to ensure both cell survival and functionality. To enhance oxygen transport, we utilized perfluorocarbon (PFC) oxygen vectors, specifically perfluorooctyl bromide (PFOB) immobilized in an alginate matrix. Metabolic activity of HepG2 liver cells encapsulated in 1% alginate/10% PFOB composite system was 47-104% higher than alginate systems lacking PFOB. A cubic model was developed to understand the oxygen transport mechanism in the alginate/PFOB composite system. The theoretical flux enhancement in alginate systems containing 10% PFOB was 18% higher than in alginate-only systems. Oxygen uptake rates (OURs) of HepG2 cells were enhanced with 10% PFOB addition under both 20% and 5% O2 boundary conditions, by 8% and 15%, respectively. Model predictions were qualitatively and quantitatively verified with direct experimental OUR measurements using both a perfusion reactor and oxygen sensing plate, demonstrating a greater OUR enhancement under physiological O2 boundary conditions (i.e., 5% O2). Inclusion of PFCs in an encapsulation matrix is a useful strategy for overcoming oxygen limitations and ensuring cell viability and functionality both for large devices (>1 mm) and over extended time periods. Although our results specifically indicate positive enhancements in metabolic activity using the model HepG2 liver system encapsulated in alginate, PFCs could be useful for improving/stabilizing oxygen supply in a wide range of cell types and hydrogels.
细胞封装为细胞提供了三维结构,以模拟生理条件,并与单层培养相比改善细胞信号传导、增殖和组织构建。封装装置常常面临传质不佳的问题,尤其是对于氧气而言,必须满足关键的溶解水平以确保细胞存活和功能。为了增强氧气传输,我们利用了全氟碳(PFC)氧载体,特别是固定在藻酸盐基质中的全氟辛基溴(PFOB)。封装在1%藻酸盐/10% PFOB复合系统中的HepG2肝细胞的代谢活性比缺乏PFOB的藻酸盐系统高47 - 104%。开发了一个立方模型来理解藻酸盐/PFOB复合系统中的氧气传输机制。含有10% PFOB的藻酸盐系统中的理论通量增强比仅含藻酸盐的系统高18%。在20%和5% O₂边界条件下,添加10% PFOB均可提高HepG2细胞的氧气摄取率(OURs),分别提高了8%和15%。使用灌注反应器和氧传感板通过直接实验OUR测量对模型预测进行了定性和定量验证,证明在生理O₂边界条件(即5% O₂)下OUR增强更大。在封装基质中包含PFC是克服氧气限制并确保大型装置(>1 mm)在较长时间段内细胞活力和功能的有用策略。尽管我们的结果具体表明使用封装在藻酸盐中的模型HepG2肝脏系统代谢活性有积极增强,但PFC可用于改善/稳定多种细胞类型和水凝胶中的氧气供应。