Spontaneous Formation of Highly Functional Three-Dimensional Multilayer from Human Hepatoma Hep G2 Cells Cultured on an Oxygen-Permeable Polydimethylsiloxane Membrane
Spontaneous Formation of Highly Functional Three-Dimensional Multilayer from Human Hepatoma Hep G2 Cells Cultured on an Oxygen-Permeable Polydimethylsiloxane Membrane
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DOI：
10.1089/ten.tec.2009.0042
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发表时间：
2010-04-01
影响因子：
3
通讯作者：
Sakai, Yasuyuki
中科院分区：
文献类型：
--
作者：
Evenou, Fanny;Fujii, Teruo;Sakai, Yasuyuki
当考虑高密度肝细胞培养时，向细胞充分输送氧气显得特别重要，因为它是肝细胞功能的最限制性参数之一。在这里，我们报告的影响，直接氧通过透气性聚二甲基硅氧烷膜对肝细胞培养。我们使用高度增殖的人肝癌Hep G2细胞来评估这种直接供氧的生长相关限制。由于更大的氧气供应，Hep G2细胞的增殖显着增强，导致形成厚的三维细胞多层。这是支持的氧浓度分布在附近的细胞层，通过数值模拟预测。所应用的细胞还显示出增加的功能性，如通过高白蛋白产生速率所评估的，其中基于每孔的白蛋白分泌在15天后达到在常规聚苯乙烯板中培养的细胞的50倍。厌氧糖酵解在同一时间显着减少，如所评估的减少的葡萄糖消耗和乳酸生产率。通过这种透气膜的直接氧合似乎适合于工程化功能性厚肝组织用于体内植入，以及用于基于微孔板的化学/药物筛选应用。
When considering high-density liver cell cultures, adequate delivery of oxygen to the cells appears particularly crucial since it is one of the most limiting parameters of hepatocytes' functions. Here we report on the effects of direct oxygenation through a gas-permeable polydimethylsiloxane membrane on liver-derived cell culture. We used highly proliferative human hepatoma Hep G2 cells to assess the growth-related limitation of such direct oxygen supply. As a consequence of the greater oxygen availability, the proliferation of Hep G2 cells was markedly enhanced, leading to the formation of a thick three-dimensional cellular multilayer. This was supported by the oxygen concentration profiles in the vicinity of the cell layers, predicted by numerical simulations. The applied cells also displayed an increased functionality as assessed by the high-albumin production rates, with per well-based albumin secretion reaching after 15 days 50 times that of the cells cultured in conventional polystyrene plates. Anaerobic glycolysis was in the same time significantly reduced as assessed by both the reduced glucose consumption and lactate production rates. Direct oxygenation through such a gas-permeable membrane appears suitable for engineering functional thick liver tissues for in vivo implantation, as well as for microplate-based chemical/drug screening applications.