Autocrine-Controlled Formation and Function of Tissue-Like Aggregates by Primary Hepatocytes in Micropatterned Hydrogel Arrays
Autocrine-Controlled Formation and Function of Tissue-Like Aggregates by Primary Hepatocytes in Micropatterned Hydrogel Arrays
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DOI：
10.1089/ten.tea.2010.0398
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发表时间：
2011-04-01
影响因子：
4.1
通讯作者：
Griffith, Linda G.
中科院分区：
文献类型：
--
作者：
Williams, Courtney M.;Mehta, Geeta;Griffith, Linda G.
肝脏执行多种基本功能，部分由自分泌信号调节，包括肝细胞产生的生长因子和细胞外基质（ECM）。自分泌因子的局部浓度受细胞表面受体介导的结合和扩散到局部基质之间的平衡控制，因此预计会受到细胞培养环境维度的影响。为了研究生长因子和ECM调节的自分泌信号在维持适当的原代肝细胞存活、代谢功能和极性中的作用，我们使用微图案化的半合成聚乙二醇-纤维蛋白原水凝胶创建了限定几何形状的三维培养物，以提供机械顺应性、非粘附性材料平台，其可以通过细胞分泌因子进行修饰。我们发现，在缺乏外源性肽生长因子或ECM的情况下，肝细胞保留表皮生长因子（EGF）受体配体（EGF和转化生长因子-α）和原癌间充质上皮转化因子（c-MET）配体肝细胞生长因子（HGF），沿着纤连蛋白。此外，在这种三维微环境中培养的肝细胞在10天的培养期内保持了高水平的肝脏特异性功能。α 5 β 1或EGF受体的功能阻断剂显著降低了细胞活力和功能，表明在没有其他外源性信号的情况下，这两种受体的信号传导是肝细胞体外存活和功能所必需的。
The liver carries out a variety of essential functions regulated in part by autocrine signaling, including hepatocyte-produced growth factors and extracellular matrix (ECM). The local concentrations of autocrine factors are governed by a balance between receptor-mediated binding at the cell surface and diffusion into the local matrix and are thus expected to be influenced by the dimensionality of the cell culture environment. To investigate the role of growth factor and ECM-modulated autocrine signaling in maintaining appropriate primary hepatocyte survival, metabolic functions, and polarity, we created three-dimensional cultures of defined geometry using micropatterned semisynthetic polyethylene glycol-fibrinogen hydrogels to provide a mechanically compliant, nonadhesive material platform that could be modified by cell-secreted factors. We found that in the absence of exogenous peptide growth factors or ECM, hepatocytes retain the epidermal growth factor (EGF) receptor ligands (EGF and transforming growth factor-alpha) and the proto-oncogenic mesenchymal epithelial transition factor (c-MET) ligand hepatocyte growth factor (HGF), along with fibronectin. Further, hepatocytes cultured in this three-dimensional microenvironment maintained high levels of liver-specific functions over the 10-day culture period. Function-blocking inhibitors of alpha 5 beta 1 or EGF receptor dramatically reduced cell viability and function, suggesting that signaling by both these receptors is needed for in vitro survival and function of hepatocytes in the absence of other exogenous signals.