Cultivating hepatocytes on printed arrays of HGF and BMP7 to characterize protective effects of these growth factors during in vitro alcohol injury.
Cultivating hepatocytes on printed arrays of HGF and BMP7 to characterize protective effects of these growth factors during in vitro alcohol injury.
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DOI：
10.1016/j.biomaterials.2010.04.006
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发表时间：
2010-08
期刊：
影响因子：
14
通讯作者：
Revzin, Alexander
中科院分区：
文献类型：
--
作者：
Jones, Caroline N.;Tuleuova, Nazgul;Lee, Ji Youn;Ramanculov, Erlan;Reddi, A. Hari;Zern, Mark A.;Revzin, Alexander
本研究的目的是研究生长因子(GF)阵列在酒精损伤时对肝脏的保护作用。将肝细胞生长因子和骨形态发生蛋白7与胶原(I)混合，机器人打印到标准玻片上，形成直径500μm的斑点阵列。原代大鼠肝细胞种植在阵列的顶部，形成大小与下面的蛋白质点相对应的簇。然后在100 mM乙醇中培养48h，损伤细胞阵列。与单纯胶原斑点培养的细胞相比，生长在GF斑点上的肝细胞的凋亡率较低。HGF/BMP7/胶原斑点的细胞凋亡率最低(TUNEL法估计为0.3%)，而单纯胶原点阵的细胞凋亡率最高(17.3%)。有趣的是，酒精诱导肝细胞凋亡的程度因印刷的GF浓度不同而不同。除了防止细胞凋亡，印刷的GFS还有助于在酒精损伤期间维持上皮表型，这一点从HGF保护的肝细胞中更高水平的E-钙粘素表达得到了证明。重要的是，生长因子微阵列可以用来研究肝脏损伤情况下的异型相互作用。为了突出这一点，将星状细胞-参与纤维化的非实质肝细胞-添加到位于HGF/胶原或仅胶原斑点阵列上的肝细胞中。将这些共培养物暴露在乙醇中，然后进行RT-PCR分析，结果显示，与对照组相比，与HGF保护的肝细胞共存的星状细胞的激活程度(纤维化程度)显著降低。总体而言，我们的结果表明，GF微阵列形式可以用于筛选生长因子的抗纤维化和抗凋亡作用，以及研究传递给特定细胞类型的信号如何调节异型细胞相互作用。
The goal of the present study was to investigate hepato-protective effects of growth factor (GF) arrays during alcohol injury. Hepatocyte growth factor (HGF) and bone morphogenetic protein (BMP)7 were mixed with collagen (I) and robotically printed onto standard glass slides to create arrays of 500 μm diameter spots. Primary rat hepatocytes were seeded on top of the arrays forming clusters corresponding in size to the underlying protein spots. Cell arrays were then injured in culture by exposure to 100 mM ethanol for 48h. Hepatocytes residing on GF spots were found to have less apoptosis then cells cultured on collagen-only spots. Least apoptosis (0.3 % as estimated by TUNEL assay) was observed on HGF/BMP7/collagen spots whereas most apoptosis (17.3%) was seen on collagen-only arrays. Interestingly, the extent of alcohol-induced apoptosis in hepatocytes varied based on the concentration of printed GF. In addition to preventing apoptosis, printed GFs contributed to maintenance of epithelial phenotype during alcohol injury as evidenced by higher levels of E-cadherin expression in HGF-protected hepatocytes. Importantly, GF microarrays could be used to investigate heterotypic interactions in the context of liver injury. To highlight this, stellate cells - nonparenchymal liver cells involved in fibrosis - were added to hepatocytes residing on arrays of either HGF/collagen or collagen-only spots. Exposure of these cocultures to ethanol followed by RT-PCR analysis revealed that stellate cells residing alongside HGF-protected hepatocytes were significantly less activated (less fibrotic) compared to controls. Overall, our results demonstrate that GF microarray format can be used to screen anti-fibrotic and anti-apoptotic effects of growth factors as well as to investigate how signals delivered to a specific cell type modulate heterotypic cellular interactions.
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DOI：
10.1021/la803635r
发表时间：
2009-04-09
期刊：
Langmuir : the ACS journal of surfaces and colloids
影响因子：
--
作者：
Lee JY;Shah SS;Yan J;Howland MC;Parikh AN;Pan T;Revzin A
通讯作者：
Revzin A
影响因子：
3.7
作者：
KATO, S;ISHII, T;AKAMATSU, N
通讯作者：
AKAMATSU, N
影响因子：
14
作者：
Marcantonio, Nicholas A.;Boehm, Cynthia A.;Griffith, Linda G.
通讯作者：
Griffith, Linda G.
影响因子：
13.5
作者：
Nakamura, T;Sakata, R;Ueno, H
通讯作者：
Ueno, H
影响因子：
4.8
作者：
Kaimori, Aki;Potter, James;Koteish, Ayman
通讯作者：
Koteish, Ayman