Microfluidic co-cultures of retinal pigment epithelial cells and vascular endothelial cells to investigate choroidal angiogenesis.
Microfluidic co-cultures of retinal pigment epithelial cells and vascular endothelial cells to investigate choroidal angiogenesis.
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DOI：
10.1038/s41598-017-03788-5
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发表时间：
2017-06-14
影响因子：
4.6
通讯作者：
Kaji H
中科院分区：
文献类型：
--
作者：
Chen LJ;Ito S;Kai H;Nagamine K;Nagai N;Nishizawa M;Abe T;Kaji H
血管生成在包括黄斑变性在内的许多疾病中起关键作用。目前，病理机制尚不清楚，缺乏合适的模型来解剖血管生成过程的调控。我们提出了一种体外血管生成过程，并通过检查人视网膜色素上皮细胞（ARPE-19）和人脐静脉内皮细胞（HUVEC）内的微流控装置的共培养进行测试。根据APRE-19单培养物的表征，在微流体装置中培养的细胞上发现紧密连接蛋白（ZO-1），但培养基条件的变化不影响渗透性试验中发现的单层的完整性。血管内皮生长因子（VEGF）分泌升高，在低血糖和缺氧条件下相比，控制。在确认HUVEC的血管生成能力后，通过将ARPE-19细胞暴露于氯化钴（II）（CoCl 2）来分析在降低的葡萄糖培养基和化学缺氧下的细胞-细胞相互作用。观察到ARPE-19和HUVEC之间的异型相互作用，但一旦ARPE-19单层开始分解，HUVEC的增殖就会受到阻碍。上述特征表明，由化学缺氧诱导的葡萄糖浓度和/或氧水平的改变导致ARPE-19中产生的VEGF升高，这反过来影响HUVEC的定向生长。
Angiogenesis plays a critical role in many diseases, including macular degeneration. At present, the pathological mechanisms remain unclear while appropriate models dissecting regulation of angiogenic processes are lacking. We propose an in vitro angiogenesis process and test it by examining the co-culture of human retinal pigmental epithelial cells (ARPE-19) and human umbilical vein endothelial cells (HUVEC) inside a microfluidic device. From characterisation of the APRE-19 monoculture, the tight junction protein (ZO-1) was found on the cells cultured in the microfluidic device but changes in the medium conditions did not affect the integrity of monolayers found in the permeability tests. Vascular endothelial growth factor (VEGF) secretion was elevated under low glucose and hypoxia conditions compared to the control. After confirming the angiogenic ability of HUVEC, the cell-cell interactions were analyzed under lowered glucose medium and chemical hypoxia by exposing ARPE-19 cells to cobalt (II) chloride (CoCl2). Heterotypic interactions between ARPE-19 and HUVEC were observed, but proliferation of HUVEC was hindered once the monolayer of ARPE-19 started breaking down. The above characterisations showed that alterations in glucose concentration and/or oxygen level as induced by chemical hypoxia causes elevations in VEGF produced in ARPE-19 which in turn affected directional growth of HUVEC.