Hendriks H F, Verhoofstad W A, Brouwer A, de Leeuw A M, Knook D L
Exp Cell Res. 1985 Sep;160(1):138-49. doi: 10.1016/0014-4827(85)90243-5.
Highly purified sinusoidal (fat-storing, Kupffer and endothelial cells) and parenchymal cells were isolated to assess the cellular distribution of vitamin A in liver of adult vitamin A-sufficient rats. A modified simple procedure was developed for the purification of fat-storing cells from rat liver. This was achieved by a single centrifugation step in a two-layer density Nycodenz gradient. Endothelial and Kupffer cells were obtained from the same gradient and further purified by centrifugal elutriation. Reverse-phase HPLC analysis showed that fat-storing cells contained about 300-fold the amount of retinyl esters present in parenchymal cells on a mg cell protein basis. In fat-storing cells, the same retinyl esters, viz. retinyl palmitate, retinyl stearate and retinyl oleate, were present as in whole liver. It was also observed that, within 12 h after intravenous injection of chylomicron [3H]retinyl ester, most of the radioactivity had accumulated in the fat-storing cells. It is concluded that fat-storing cells are the main storage sites for vitamin A in rat liver.
分离出高度纯化的肝血窦细胞（贮脂细胞、库普弗细胞和内皮细胞）和实质细胞，以评估成年维生素A充足大鼠肝脏中维生素A的细胞分布。开发了一种改良的简单方法，用于从大鼠肝脏中纯化贮脂细胞。这通过在两层密度的Nycodenz梯度中进行单次离心步骤来实现。内皮细胞和库普弗细胞从同一梯度中获得，并通过离心淘析进一步纯化。反相高效液相色谱分析表明，以毫克细胞蛋白计，贮脂细胞中视黄酯的含量约为实质细胞中的300倍。在贮脂细胞中，与全肝中相同的视黄酯，即棕榈酸视黄酯、硬脂酸视黄酯和油酸视黄酯存在。还观察到，静脉注射乳糜微粒[3H]视黄酯后12小时内，大部分放射性已积聚在贮脂细胞中。得出的结论是，贮脂细胞是大鼠肝脏中维生素A的主要储存部位。
