Kreamer B L, Staecker J L, Sawada N, Sattler G L, Hsia M T, Pitot H C
In Vitro Cell Dev Biol. 1986 Apr;22(4):201-11. doi: 10.1007/BF02623304.
A simple yet effective method (iso-density percoll centrifugation) has been developed for consistently preparing isolated rat liver parenchymal cells with over 98% initial viability. The method has been applied to cells isolated by a variety of collagenase digestion techniques. This procedure involves the low-speed centrifugation (50 X g) of the initial cell suspension through a percoll medium having a density of 1.06 g/ml and results in the separation of single and viable parenchymal cells from cell aggregates, debris, and nonparenchymal cells. The enriched parenchymal cells have been shown to be superior to untreated cells by a number of criteria including: preparation homogeneity, cell morphology, maintenance of cytochrome P-450, hormonal responsiveness (measured by the induction of tyrosine aminotransferase after treatment with glucagon or dexamethasone, or both), plasma membrane integrity (determined by both trypan blue exclusion and leakage of glutamic-oxaloacetic transaminase), and the DNA repair capability after treatment with benzo[a]pyrene or 2-acetylaminofluorene.
已经开发出一种简单而有效的方法（等密度 Percoll 离心法），用于持续制备初始活力超过 98%的分离大鼠肝实质细胞。该方法已应用于通过多种胶原酶消化技术分离的细胞。此程序包括将初始细胞悬液以低速（50×g）通过密度为 1.06 g/ml 的 Percoll 介质进行离心，从而从细胞聚集体、碎片和非实质细胞中分离出单个且有活力的实质细胞。通过许多标准表明，富集的实质细胞优于未处理的细胞，这些标准包括：制备均匀性、细胞形态、细胞色素 P - 450 的维持、激素反应性（通过用胰高血糖素或地塞米松或两者处理后酪氨酸转氨酶的诱导来测量）、质膜完整性（通过台盼蓝排斥和谷氨酸 - 草酰乙酸转氨酶的泄漏来确定）以及用苯并[a]芘或 2 - 乙酰氨基芴处理后的 DNA 修复能力。
