A rapid method for the isolation of large quantities of rat liver parenchymal cells with high anabolic rates.
A rapid method for the isolation of large quantities of rat liver parenchymal cells with high anabolic rates.
复制标题
一种快速分离大量具有高合成代谢率的大鼠肝实质细胞的方法。
DOI：
10.1016/0006-291x(72)90728-0
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发表时间：
1972
影响因子：
3.1
通讯作者：
S. R. Wagle
中科院分区：
文献类型：
--
作者：
W. R. Ingebretsen;S. R. Wagle
介绍了一种体外肝灌注快速分离大鼠肝实质细胞的方法。从肝切除术到细胞分离，暴露于胶原酶（20 mg/100 ml） 10-15分钟，该过程需要40-50分钟。在最佳条件下，从200 g大鼠肝脏中分离4.5 ~ 6.0× 10 8个细胞（3.0 g细胞）。用这种方法分离的细胞中，有90%到95%的细胞排除了重要的染色，在光镜下观察时没有损伤。在乳酸、丙氨酸和丙酮酸存在的情况下，这些细胞积极地将NaHC 14o 3合并到葡萄糖和蛋白质中。糖异生前体产生的葡萄糖与在离体灌注肝脏中观察到的相同。与肝脏切片或离体灌注的肝脏不同，这些细胞的糖异生率随着乳酸浓度在20 mM以上的增加而增加。80 mM的乳酸浓度刺激葡萄糖产生至体内计算的糖异生率的80 - 90%。
A rapid method for the isolation of rat liver parenchymal cells using in vitro liver perfusion is described. The procedure requires 40–50 min from hepatectomy to cell isolation with exposure to collagenase (20 mg/100 ml) for 10–15 min. Under optimal conditions 4.5 to 6.0× 10 8 cells (3.0 g of cells) were isolated from the liver of a 200 g rat. Ninety to 95% of the cells isolated by this method excluded a vital stain and were undamaged when viewed with light microscopy. These cells actively incorporate NaHC 14 O 3 into glucose and protein in the presence of lactate, alanine and pyruvate. Glucose production from gluconeogenic precursors was equivalent to that observed with the isolated perfused liver. Unlike liver slices or the isolated perfused liver, these cells show increasing gluconeogenic rates with increasing lactate concentrations above 20 mM. A concentration of 80 mM stimulated glucose production to 80–90% of the calculated in vivo gluconeogenic rate.