Basic Research
Copyright ©2006 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Gastroenterol. May 21, 2006; 12(19): 3050-3054
Published online May 21, 2006. doi: 10.3748/wjg.v12.i19.3050
An efficient method of sorting liver stem cells by using immuno-magnetic microbeads
Yu-Fei He, Yin-Kun Liu, Dong-Mei Gao, Jun Chen, Peng-Yuan Yang
Yu-Fei He, Yin-Kun Liu, Dong-Mei Gao, Jun Chen, Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai 200032, China
Yu-Fei He, Peng-Yuan Yang, Center of Proteomics, BioMedical Institutes, Fudan University, Shanghai 200032, China
Author contributions: All authors contributed equally to the work
Supported by Shanghai Science and Technology Development Program, No. 03DZ14024; the National High Technology 863 Programs, No. 2002BAC11A11 and National Development Program (973) for Key Basic Research of China, No. 2001CB510205
Correspondence to: Professor Yin-Kun Liu, Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai 200032, China. ykliu@zshospital.com
Telephone: +86-21-64041990-2501 Fax: +86-21-64037181
Received: November 11, 2005
Revised: December 2, 2005
Accepted: December 7, 2005
Published online: May 21, 2006
Abstract

AIM: To develop a method to isolate liver stem cells fast and efficiently.

METHODS: Fetal mouse liver cells were characterized by cell surface antigens (c-Kit and CD45/TER119) using flow cytometry. The candidate liver stem cells were sorted by using immuno-magnetic microbeads and identified by clone-forming culture, RT-PCR and immunofluorescence assays.

RESULTS: The c-Kit–(CD45/TER119)– cell population with 97.9% of purity were purified by immuno-magnetic microbeads at one time. The yield of this separation was about 6% of the total sorting cells and the cell viability was above 98%. When cultured in vitro these cells had high clone-forming and self-renewing ability and expressed markers of hepatocytes and bile duct cells. Functionally mature hepatocytes were observed after 21 d of culture.

CONCLUSION: This method offers an excellent tool for the enrichment of liver stem cells with high purity and viability, which could be used for further studies. It is fast, efficient, simple and not expensive.

Keywords: Liver stem cells; Immuno-magnetic microbeads; Sorting; Flow cytometry