Rotem A, Toner M, Bhatia S, Foy B D, Tompkins R G, Yarmush M L
Surgical Services, Massachusetts General Hospital, and the Shriners Burns Institute, Department of Surgery, Harvard Medical School, Boston, Massachusetts 02114.
Biotechnol Bioeng. 1994 Mar 25;43(7):654-60. doi: 10.1002/bit.260430715.
Many recent studies related to the development of bioartificial liver devices have utilized hepatocytes cultured within devices of various geometries. Because hepatocytes are anchorage-dependent cells, they need to attach and spread onto the extracellular matrix to be able to function, a process that requires energy. Thus, it is important to deliver enough oxygen to hepatocytes contained within bioartificial liver devices during the early phase of cellular organization while the cells interact with the extracellular matrix. In this study, we investigated the effect of oxygen on the attachment and spreading of hepatocytes. Increasing the gas phase oxygen from 0 to 160 mmHg resulted in an increase in the percentage of cells attaching from 43.0 +/- 5.8% to 103.6 +/- 29%, 1 h after seeding. In a similar manner, increasing the gas phase oxygen from 0 to 160 mmHg resulted in an increase of the projected surface area from 310 +/- 35 to 827 +/- 127 mum(2), 24 h after seeding. Furthermore, the partial pressure of oxygen at the cell level was estimated using a diffusion-reaction model. The model indicated that a cell surface oxygen partial pressure of 0.064 mmHg was required for the half-maximal (K(m) (a)) attachment of hepatocytes to collagen-based substrate. On the other hand, the K(m) (s) value of the spreading process was predicted to be 0.13 mmHg. The results of this study demonstrate the importance of oxygen during the initial stages of attachment and spreading of hepatocytes, and it has important implications in the design of hepatocyte-based bioartificial liver devices. (c) 1994 John Wiley & Sons, Inc.
近期许多与生物人工肝装置研发相关的研究都利用了在各种几何形状装置中培养的肝细胞。由于肝细胞是贴壁依赖性细胞，它们需要附着并铺展在细胞外基质上才能发挥功能，而这一过程需要能量。因此，在细胞与细胞外基质相互作用的细胞组织早期阶段，向生物人工肝装置内的肝细胞输送足够的氧气非常重要。在本研究中，我们调查了氧气对肝细胞附着和铺展的影响。接种1小时后，将气相氧气从0 mmHg增加到160 mmHg，导致附着细胞的百分比从43.0±5.8%增加到103.6±29%。以类似的方式，接种24小时后，将气相氧气从0 mmHg增加到160 mmHg，导致投影表面积从310±35增加到827±127μm²。此外，使用扩散反应模型估计了细胞水平的氧分压。该模型表明，肝细胞与基于胶原蛋白的底物半最大附着（K(m)(a)）所需的细胞表面氧分压为0.064 mmHg。另一方面，铺展过程的K(m)(s)值预计为0.13 mmHg。本研究结果证明了氧气在肝细胞附着和铺展初始阶段的重要性，并且对基于肝细胞的生物人工肝装置的设计具有重要意义。(c) 1994 John Wiley & Sons, Inc.
