A New Fluidized Bed Bioreactor Based on Diversion-Type Microcapsule Suspension for Bioartificial Liver Systems.
A New Fluidized Bed Bioreactor Based on Diversion-Type Microcapsule Suspension for Bioartificial Liver Systems.
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DOI：
10.1371/journal.pone.0147376
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Li L
中科院分区：
文献类型：
--
作者：
Lu J;Zhang X;Li J;Yu L;Chen E;Zhu D;Zhang Y;Li L
含有包封肝细胞的流化床生物反应器可能是中空纤维生物反应器的一个有价值的替代品，可以实现改善的传质和扩大临床使用所需的潜力。然而，传统的流化床生物反应器（FBB）在高灌注速度下运行，由于其对含有细胞的微胶囊的破坏和大空隙体积而无法提供所需的性能。在这项研究中，我们开发了一种新型的导流型微胶囊悬浮流化床生物反应器（DMFBB）。考察了海藻酸盐/壳聚糖微胶囊在不同流速下的生物反应器空隙体积和稳定性。测定包膜肝细胞系（C3A细胞）的细胞活力、合成和代谢功能以及转录水平上代谢酶的表达。新型生物反应器的空隙体积明显小于传统流化床反应器。此外，微胶囊在DMFBB流化过程中受到的破坏较小，这可以从微胶囊保留率、肿胀和破裂的结果中得到反映。包裹的C3A细胞在DMFBB中比在FBB中表现出更高的活力和CYP1A2和CYP3A4活性，尽管白蛋白和尿素合成的增加不那么明显。DMFBB细胞中几个cyp450相关基因和一个白蛋白相关基因的转录水平显著高于FBB细胞。综上所述，我们的研究结果表明，DMFBB是基于包封肝细胞的流化床生物反应器设计生物人工肝系统的一个有希望的替代方案，可用于治疗急性肝功能衰竭或其他严重肝脏疾病的患者。
A fluidized bed bioreactor containing encapsulated hepatocytes may be a valuable alternative to a hollow fiber bioreactor for achieving the improved mass transfer and scale-up potential necessary for clinical use. However, a conventional fluidized bed bioreactor (FBB) operating under high perfusion velocity is incapable of providing the desired performance due to the resulting damage to cell-containing microcapsules and large void volume. In this study, we developed a novel diversion-type microcapsule-suspension fluidized bed bioreactor (DMFBB). The void volume in the bioreactor and stability of alginate/chitosan microcapsules were investigated under different flow rates. Cell viability, synthesis and metabolism functions, and expression of metabolizing enzymes at transcriptional levels in an encapsulated hepatocyte line (C3A cells) were determined. The void volume was significantly less in the novel bioreactor than in the conventional FBB. In addition, the microcapsules were less damaged in the DMFBB during the fluidization process as reflected by the results for microcapsule retention rates, swelling, and breakage. Encapsulated C3A cells exhibited greater viability and CYP1A2 and CYP3A4 activity in the DMFBB than in the FBB, although the increases in albumin and urea synthesis were less prominent. The transcription levels of several CYP450-related genes and an albumin-related gene were dramatically greater in cells in the DMFBB than in those in the FBB. Taken together, our results suggest that the DMFBB is a promising alternative for the design of a bioartificial liver system based on a fluidized bed bioreactor with encapsulated hepatocytes for treating patients with acute hepatic failure or other severe liver diseases.