Layered long-term co-culture of hepatocytes and endothelial cells on a transwell membrane: toward engineering the liver sinusoid.
Layered long-term co-culture of hepatocytes and endothelial cells on a transwell membrane: toward engineering the liver sinusoid.
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DOI：
10.1088/1758-5082/5/4/045008
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发表时间：
2013-12
期刊：
影响因子：
9
通讯作者：
Noh HM
中科院分区：
文献类型：
--
作者：
Kang YB;Rawat S;Cirillo J;Bouchard M;Noh HM
本文提出了一种新的肝脏模型，模拟肝窦，大多数肝脏活动发生。我们目前的肝脏模型的一个关键方面是原代大鼠肝细胞（PRH）和原代大鼠肝窦内皮细胞（LSEC）或牛主动脉内皮细胞（BAEC）在transwell膜上的分层共培养。当尝试分层共培养时，在肝细胞和内皮细胞之间放置薄的基质胶层以模拟Disse间隙，细胞没有形成完全分离的单层。然而，当肝细胞和内皮细胞培养的transwell膜的相对侧，PRHs与LSEC或BAECs共培养保持其活力和正常形态分别为39和57天。我们评估了肝细胞特异性分化标志物的存在，以验证PRH在长期共培养中保持分化，并通过监测尿素合成来分析肝细胞功能。我们还注意到，从第13天到第48天，细胞色素P-450的表达在共培养系统中保持相似。因此，我们的新的肝脏模型系统表明，原代肝细胞可以培养延长的时间，并保留其肝细胞特异性功能时，与内皮细胞分层。
This paper presents a novel liver model that mimics the liver sinusoid where most liver activities occur. A key aspect of our current liver model is a layered co-culture of primary rat hepatocytes (PRHs) and primary rat liver sinusoidal endothelial cells (LSECs) or bovine aortic endothelial cells (BAECs) on a transwell membrane. When a layered co-culture was attempted with a thin matrigel layer placed between hepatocytes and endothelial cells to mimic the Space of Disse, the cells did not form completely separated monolayers. However, when hepatocytes and endothelial cells were cultured on the opposite sides of a transwell membrane, PRHs co-cultured with LSECs or BAECs maintained their viability and normal morphology for 39 and 57 days, respectively. We assessed the presence of hepatocyte-specific differentiation markers to verify that PRHs remained differentiated in the long-term co-culture and analyzed hepatocyte function by monitoring urea synthesis. We also noted that the expression of cytochrome P-450 remained similar in the co-cultured system from Day 13 to Day 48. Thus, our novel liver model system demonstrated that primary hepatocytes can be cultured for extended times and retain their hepatocyte-specific functions when layered with endothelial cells.
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影响因子：
1.8
作者：
Doi, K;Hargitai, J;Gouras, P
通讯作者：
Gouras, P
DOI：
10.1007/10_029
发表时间：
2007-01-01
期刊：
TISSUE ENGINEERING II: BASICS OF TISSUE ENGINEERING AND TISSUE APPLICATIONS
影响因子：
--
作者：
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通讯作者：
Yarmush, Martin L.
影响因子：
3.8
作者：
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通讯作者：
Griffith, LG
影响因子：
14
作者：
Chen, Alice A.;Khetani, Salman R.;Lee, Sunyoung;Bhatia, Sangeeta N.;Van Vliet, Krystyn J.
通讯作者：
Van Vliet, Krystyn J.
影响因子：
8
作者：
Huang, Jianmin;Karakucuk, Vildan;Rhoads, David B.
通讯作者：
Rhoads, David B.