An efficient method of sorting liver stem cells by using immuno-magnetic microbeads.
OBJECTIVE
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.


CONCLUSIONS
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.
He
Yu-Fei
YF
Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
Liu
Yin-Kun
YK
Gao
Dong-Mei
DM
Chen
Jun
J
Yang
Peng-Yuan
PY
D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D019070:Cell Lineage; D002478:Cells, Cultured; D005260:Female; D005434:Flow Cytometry; D018189:Immunomagnetic Separation / Q000379:methods*; D017493:Leukocyte Common Antigens / Q000032:analysis; D008099:Liver / Q000166:cytology* / Q000196:embryology* / Q000276:immunology / Q000502:physiology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D019009:Proto-Oncogene Proteins c-kit / Q000032:analysis; D013234:Stem Cells / Q000166:cytology* / Q000276:immunology / Q000502:physiology
D019009:Proto-Oncogene Proteins c-kit; D017493:Leukocyte Common Antigens