Evaluation of a bioreactor with stacked sheet shaped organoids of primary hepatocytes
Evaluation of a bioreactor with stacked sheet shaped organoids of primary hepatocytes
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DOI：
10.1016/j.jbiosc.2009.01.005
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发表时间：
2009-05-01
影响因子：
2.8
通讯作者：
Kajiwara, Toshihisa
中科院分区：
文献类型：
--
作者：
Kusumi, Tomoaki;Ishihara, Kazuhisa;Kajiwara, Toshihisa
肝细胞类器官具有体内样细胞形态，并在体外维持细胞活力和功能。另一方面，对肝细胞的氧供应有时在厚度超过100 μ m的类器官的核心中受到限制。在这项研究中，我们设计和检查了一种新的生物反应器，使用片状类器官（类器官片），其中厚度受到控制，以防止由于氧气供应的限制而导致的类器官核心中的肝细胞死亡。细胞培养空间由堆叠的类器官形成空间和培养基流动通道组成。每个空间由平的多孔聚碳酸酯膜隔开，并且用不锈钢间隔物将类器官厚度控制在100 μ m。将新鲜分离的肝细胞（7.0 x 10（7））固定在生物反应器中，产生的细胞密度为4.5 x 10（7）个细胞/cm（3）-生物反应器。在测试的五种流速（1.0、5.0、10、20和50 mL/min）中，具有10 mL/min的生物反应器具有最高的氨去除和白蛋白分泌活性，持续至少14天。总之，控制类器官厚度的新生物反应器可用于实现高细胞密度培养和维持肝细胞功能，以避免由于氧气供应限制而导致类器官核心中的细胞死亡。该生物反应器可用于开发使用培养的肝细胞的各种应用。(c)2009年，生物技术学会，日本。All rights reserved.
Hepatocyte organoids have an in vivo-like cell morphology and maintain cell viability and function in vitro. On the other hand, the oxygen supply to hepatocytes is sometimes limited in the core of organoids that are more than 100 pm in thickness. In this study, we designed and examined a new bioreactor using sheet-shaped organoids (organoid-sheets) in which the thickness was controlled to prevent hepatocyte death in the core of organoid due to limitation of oxygen supply. The cell culture space consisted of stacked organoid formation spaces and medium flow channels. Each space was separated by flat porous polycarbonate membranes, and the organoid thickness was controlled at 100 mu m with a stainless steel spacer. Freshly isolated hepatocytes (7.0 x 10(7)) were immobilized in the bioreactor, yielding a cell density of 4.5 x 10(7) cells/cm(3)-bioreactor. Of the five flow rates tested (1.0, 5.0, 10, 20, and 50 mL/min), the bioreactor with the 10 mL/min had the highest ammonia removal and albumin secretion activities for at least 14 days. In conclusion, a new bioreactor controlling organoid thickness is useful for achieving high cell density culture and the maintenance of hepatocyte function to avoid cell death in the core of the organoids due to limitation of oxygen supply. The bioreactor may be useful for the development of various applications using cultured hepatocytes. (c) 2009, The Society for Biotechnology, Japan. All rights reserved.