Alpini G, Phillips J O, Vroman B, LaRusso N F
Center for Basic Research in Digestive Diseases, Mayo Medical School, Clinic and Foundation, Rochester, Minnesota 55905.
Hepatology. 1994 Aug;20(2):494-514.
The development of new and refined separation techniques--including FACS, FFE, CFE and isopyknic gradients--has had a profound impact on the ability of investigators to isolate specific cell types from the liver. Although some of these techniques, such as FFE, may be of limited preparative value, they are nonetheless important analytical tools that detect subtle differences among cell subpopulations. The isolation of highly purified preparations of liver cells in large yields requires the use of more conventional purification methods such as CFE and isopyknic centrifugation. Immunological approaches represent a key development for the isolation of specific liver cell types, especially when they are used in combination with other techniques. Excellent, reliable and relatively simple techniques now exist to isolate highly purified preparations of hepatocytes, cholangiocytes, KCs, SCs, FSC, myofibroblasts and pit cells. Additional work is necessary to refine techniques for the isolation of dendritic cells and lymphocytes.
新型精细分离技术的发展——包括荧光激活细胞分选术(FACS)、自由流电泳(FFE)、连续流离心(CFE)和等密度梯度离心——对研究人员从肝脏中分离特定细胞类型的能力产生了深远影响。尽管其中一些技术,如FFE,可能在制备方面价值有限,但它们仍然是检测细胞亚群间细微差异的重要分析工具。要大量获得高度纯化的肝细胞制剂,需要使用更传统的纯化方法,如CFE和等密度离心。免疫方法是分离特定肝细胞类型的关键进展,尤其是当它们与其他技术结合使用时。现在已有出色、可靠且相对简单的技术来分离高度纯化的肝细胞、胆管细胞、库普弗细胞(KCs)、肝星状细胞(SCs)、纤维状星形细胞(FSC)、肌成纤维细胞和陷窝细胞制剂。还需要进一步开展工作来完善分离树突状细胞和淋巴细胞的技术。