期刊
PROTEOMICS
卷 11, 期 17, 页码 3556-3564出版社
WILEY-BLACKWELL
DOI: 10.1002/pmic.201100157
关键词
Hepatic stellate cells; Hepatocytes; Kupffer cells; Liver sinusoidal endothelial cells; Magnetic activated cell sorting; Technology
资金
- Chinese State Key Projects for Basic Research [2011CB910601, 2011CB910700, 2010CB912700, 2011CB505304]
- National Natural Science Foundation of China [30972909, 81000192, 81001470, 81010064]
- International Scientific Collaboration Program [2009DFB33070, 2010DFA31260, 2011DFB30370]
- State Key Laboratory of Proteomics [SKLP-Y200901, SKLP-O200901]
It becomes increasingly clear that separation of pure cell populations provides a uniquely sensitive and accurate approach to protein profiling in biological systems and opens up a new area for proteomic analysis. The method we described could simultaneously isolate population of hepatocytes (HCs), hepatic stellate cells (HSCs), Kupffer cells (KCs) and liver sinusoidal endothelial cells (LSECs) by a combination of collagenase-based density gradient centrifugation and magnetic activated cell sorting with high purity and yield for the first time. More than 98% of the isolated HCs were positive for cytokeratin 18, with a viability of 91%. Approximately 97% of the isolated HSCs expressed glial fibrillary acidic protein with a viability of 95%. Nearly 98% of isolated KCs expressed F4/80 with a viability of 94%. And the purity of LSECs reached up to 91% with a viability of 94%. And yield for HCs, HSCs, LSECs and KCs were 6.3, 1.3, 2.6 and 5.0 million per mouse. This systematic isolation method enables us to study the proteome profiling of different types of liver cells with high purity and yield, which is especially useful for sample preparation of Human Liver Proteome Project.
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