3D hepatic cultures simultaneously maintain primary hepatocyte and liver sinusoidal endothelial cell phenotypes.
3D hepatic cultures simultaneously maintain primary hepatocyte and liver sinusoidal endothelial cell phenotypes.
复制标题
3D 肝培养同时维持原代肝细胞和肝窦内皮细胞表型。
DOI：
10.1371/journal.pone.0015456
复制
发表时间：
2010-11-12
期刊：
影响因子：
3.7
通讯作者：
Rajagopalan P
中科院分区：
文献类型：
--
作者：
Kim Y;Rajagopalan P
开发表现出稳定表型的体外工程肝组织是肝组织工程领域的主要挑战。然而，当肝实质（肝细胞）和非实质（肝窦内皮细胞，LSEC）细胞类型从体内自然环境中去除时，其快速去分化仍然是一个主要障碍。这项研究的主要目标是证明以分层结构培养的肝细胞可以保留或潜在增强两种细胞类型的肝脏特异性行为。原代大鼠肝细胞和大鼠 LSEC (rLSEC) 以分层三维 (3D) 结构培养。细胞层由壳聚糖-透明质酸聚电解质多层 (PEM) 分隔，用于模拟迪斯空间。肝细胞和 rLSEC 在 12 天的培养期内表现出几个关键的表型特征。正弦内皮 1 抗体 (SE-1) 的免疫染色表明，在 3D 肝模型中培养的 rLSEC 在 12 天的时间内保持了这种独特的特征。相比之下，单层培养的 rLSEC 在三天内就失去了表型。 3D 培养物独特的分层结构增强了异型细胞间的相互作用，从而改善了肝细胞功能。 rLSEC-PEM-肝细胞培养物中的白蛋白产量增加了三到六倍。仅 rLSEC-PEM-肝细胞培养物表现出 CYP1A1/2 和 CYP3A 活性增加。仅在 rLSEC-PEM-肝细胞培养物中观察到明确的胆小管。总之，这些数据表明 rLSEC-PEM-肝细胞培养物非常适合监测肝脏中毒素的转化及其从该器官的转运。总之，这些结果表明，分层的 rLSEC-PEM-肝细胞模型概括了肝窦的关键特征，是获得体外肝脏代谢、解毒和信号通路综合知识的潜在强大媒介。
Developing in vitro engineered hepatic tissues that exhibit stable phenotype is a major challenge in the field of hepatic tissue engineering. However, the rapid dedifferentiation of hepatic parenchymal (hepatocytes) and non-parenchymal (liver sinusoidal endothelial, LSEC) cell types when removed from their natural environment in vivo remains a major obstacle. The primary goal of this study was to demonstrate that hepatic cells cultured in layered architectures could preserve or potentially enhance liver-specific behavior of both cell types. Primary rat hepatocytes and rat LSECs (rLSECs) were cultured in a layered three-dimensional (3D) configuration. The cell layers were separated by a chitosan-hyaluronic acid polyelectrolyte multilayer (PEM), which served to mimic the Space of Disse. Hepatocytes and rLSECs exhibited several key phenotypic characteristics over a twelve day culture period. Immunostaining for the sinusoidal endothelial 1 antibody (SE-1) demonstrated that rLSECs cultured in the 3D hepatic model maintained this unique feature over twelve days. In contrast, rLSECs cultured in monolayers lost their phenotype within three days. The unique stratified structure of the 3D culture resulted in enhanced heterotypic cell-cell interactions, which led to improvements in hepatocyte functions. Albumin production increased three to six fold in the rLSEC-PEM-Hepatocyte cultures. Only rLSEC-PEM-Hepatocyte cultures exhibited increasing CYP1A1/2 and CYP3A activity. Well-defined bile canaliculi were observed only in the rLSEC-PEM-Hepatocyte cultures. Together, these data suggest that rLSEC-PEM-Hepatocyte cultures are highly suitable models to monitor the transformation of toxins in the liver and their transport out of this organ. In summary, these results indicate that the layered rLSEC-PEM-hepatocyte model, which recapitulates key features of hepatic sinusoids, is a potentially powerful medium for obtaining comprehensive knowledge on liver metabolism, detoxification and signaling pathways in vitro.
登录
查看更多内容
DOI：
10.1186/1476-5926-3-4
发表时间：
2004-08-12
期刊：
Comparative hepatology
影响因子：
--
作者：
Elvevold KH;Nedredal GI;Revhaug A;Smedsrød B
通讯作者：
Smedsrød B
影响因子：
2.9
作者：
DUNN, JCY;TOMPKINS, RG;YARMUSH, ML
通讯作者：
YARMUSH, ML
影响因子：
10.8
作者：
Park, IK;Kim, TH;Cho, CS
通讯作者：
Cho, CS
影响因子：
14
作者：
Joddar, Binata;Ramamurthi, Anand
通讯作者：
Ramamurthi, Anand
影响因子：
13.5
作者：
Gao, WS;Bentley, RC;Clavien, PA
通讯作者：
Clavien, PA