TGF-β1 regulation in hepatocyte-NIH3T3 co-culture is important for the enhanced hepatocyte function in 3D microenvironment
TGF-β1 regulation in hepatocyte-NIH3T3 co-culture is important for the enhanced hepatocyte function in 3D microenvironment
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DOI：
10.1002/bit.20372
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发表时间：
2005-03-05
影响因子：
3.8
通讯作者：
Yu, H
中科院分区：
文献类型：
--
作者：
Chia, SM;Lin, PC;Yu, H
肝细胞或肝细胞球状体与支持NIH 3 T3在由混合天然/合成基质形成的3D微胶囊中的共培养导致肝细胞功能增强。我们研究了共培养中可溶性因子和直接细胞-细胞相互作用的作用机制。来自共培养物的条件培养基在微囊化肝细胞中诱导比来自NIH 3 T3-或肝细胞-单独对照的条件培养基更高的P450细胞色素氧化酶活性（由EROD测定指示）。通过膜插入物物理分离的肝细胞-NIH 3 T3共培养物的条件培养基降低了功能增强。在已知的肝细胞功能刺激物中，TGF β 1主要负责在该3D共培养物中刺激肝细胞功能，因为通过抗体消耗去除TGF β 1消除了功能增强，并且TGF β 1的重建恢复了功能增强。在细胞外环境中潜伏的TGF β 1的激活在共培养中上调，在转录和翻译水平上没有观察到TGF β 1表达的增加。我们的数据使我们更好地理解了共培养如何在体外增强肝细胞功能，并为肝组织工程、药物代谢研究和其他需要体外培养功能性肝细胞的应用的进一步创新铺平了道路。(C)2005 Wiley Periodicals，Inc.
Co-culture of hepatocytes or hepatocyte spheroids with the supporting NIH3T3 in a 3D microcapsule formed with a hybrid natural/synthetic matrix has led to enhanced hepatocyte functions. We investigated the mechanism of the functional enhancement in co-culture with respect to the contributions of soluble factors and direct cell-cell interactions. The conditioned media from the co-culture induced higher P450 cytochrome oxidase activity (indicated by EROD assay) in the microencapsulated hepatocytes than the conditioned media from the NIH3T3- or the hepatocytes-alone controls. Conditioned media from physically separated co-culture of hepatocytes-NIH3T3 by a membrane insert reduced the functional enhancement. Among the known stimulators of hepatocyte functions, TGFbeta1 is primarily responsible for the stimulation of hepatocyte functions in this 3D co-culture since the removal of TGFbeta1 by antibody depletion eliminated the functional enhancement and the reconstitution of TGFbeta1 restored the functional enhancement. Activation of latent TGFbeta1 in an extracellular environment were upregulated in co-culture with no observable increase in the TGFbeta1 expression at transcriptional and translational levels. Our data led to an improved understanding of how co-culture enhances hepatocyte functions in vitro and pave the way for further innovations in liver tissue engineering, drug metabolism studies, and other applications that require functional hepatocytes cultured in vitro. (C) 2005 Wiley Periodicals, Inc.