Transcellular transport of fluorescein in hepatocyte monolayers: evidence for functional polarity of cells in culture.
Transcellular transport of fluorescein in hepatocyte monolayers: evidence for functional polarity of cells in culture.
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肝细胞单层中荧光素的跨细胞转运：培养细胞功能极性的证据。
DOI：
10.1073/pnas.79.16.4985
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发表时间：
1982
影响因子：
11.1
通讯作者：
L. Schwarz
中科院分区：
文献类型：
--
作者：
C. Barth;L. Schwarz
活体大鼠肝脏将血液中的胆汁盐、蛋白质和染料转移到胆汁中。本通讯的目的是证明这种跨细胞转运在培养的成年大鼠肝细胞中的维持。在培养基中加入荧光素（20微克/毫升）2分钟后，荧光显微镜观察到细胞荧光达到最大值。随后，细胞间隙的荧光逐渐增强，在5 ~ 20分钟达到最大值，形成了一个明亮的细胞间隙荧光网络。以下观察结果可以作为证据，证明这些发现反映了细胞对染料的摄取和小管分泌。首先，在添加荧光素二醋酸酯（荧光素的非荧光前体）时进行相同的观察序列，证明该化合物在分泌到细胞间隙之前已在细胞中被吸收和代谢为荧光素。其次，用抑胆盐牛磺酸胆石酸盐（100 μ mol/l）对单层细胞进行预孵育，几乎完全抑制细胞间荧光，但不能抑制细胞荧光。由此得出结论，培养的肝细胞显示出一种功能性极性，允许胆汁分泌物质的跨细胞运输。
The rat liver in vivo transfers bile salts, proteins, and dyes from blood into bile. It is the purpose of this communication to demonstrate the maintenance of this transcellular transport in cultured adult rat hepatocytes. Two minutes after adding fluorescein (20 microgram/ml) to the culture medium, maximal cellular fluorescence was observed through the fluorescence microscope. Subsequently, intercellular clefts showed a steadily increasing fluorescence with a maximum between 5 and 20 min, resulting in a brightly fluorescent network of intercellular gaps. The following observations are taken as evidence that these findings reflect cellular uptake and canalicular secretion of the dye. First, the same sequence of observations was made upon addition of fluorescein diacetate (a nonfluorescent precursor of fluorescein), proving that the compound had been taken up and metabolized in the cells to fluorescein before secretion into intercellular clefts. Second, preincubation of the monolayers with the cholestatic bile salt taurolithocholate (100 mumol/liter) suppressed almost completely intercellular but not cellular fluorescence. It is concluded that hepatocytes in culture show a functional polarity permitting the transcellular transport of substances bound for biliary secretion.