Purification of fetal liver stem/progenitor cells containing all the repopulation potential for normal adult rat liver.
OBJECTIVE
Previously, we showed high-level, long-term liver replacement after transplantation of unfractionated embryonic day (ED) 14 fetal liver stem/progenitor cells (FLSPC). However, for clinical applications, it will be essential to transplant highly enriched cells, while maintaining high repopulation potential.


METHODS
Dlk-1, a member of the delta-like family of cell surface transmembrane proteins, is highly expressed in human and rodent fetal liver. Dlk-1(+) cells, isolated from ED14 fetal liver using immunomagnetic beads, were examined for their hepatic gene expression profile and characteristic properties in vitro and their proliferative and differentiation potential in vivo after transplantation into normal adult rat liver.


RESULTS
Rat ED14 FLSPC were purified to 95% homogeneity and exhibited cell culture and gene expression characteristics expected for hepatic stem/progenitor cells. Rat ED14 FLSPC are alpha-fetoprotein(+)/cytokeratin-19(+) or alpha-fetoprotein(+)/cytokeratin-19(-) and contain all of the normal liver repopulation capacity found in fetal liver. Hematopoietic stem cells, a major component in crude fetal liver cell preparations that engraft in other organs, such as bone marrow, spleen, and lung, are totally removed by Dlk-1 selection, and Dlk-1 purified FLSPC repopulate only the liver.


CONCLUSIONS
This is the first study reporting purification of hepatic stem/progenitor cells from fetal liver that are fully capable of repopulating the normal adult liver. This represents a major advance toward developing protocols that will be essential for clinical application of liver cell transplantation therapy.
Oertel
Michael
M
Marion Bessin Liver Research Center, Albert Einstein College of Medicine of Yeshiva University, Bronx, New York, USA.
Menthena
Anuradha
A
Chen
Yuan-Qing
YQ
Teisner
Børge
B
Jensen
Charlotte Harken
CH
Shafritz
David A
DA
D000818:Animals; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D018819:Dipeptidyl Peptidase 4 / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000201:enzymology / Q000378:metabolism / Q000637:transplantation*; D005786:Gene Expression Regulation; D006498:Hepatectomy; D022781:Hepatocytes / Q000201:enzymology / Q000378:metabolism / Q000637:transplantation*; D018189:Immunomagnetic Separation; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D008099:Liver / Q000166:cytology / Q000196:embryology / Q000378:metabolism / Q000601:surgery*; D008115:Liver Regeneration / Q000235:genetics; D008565:Membrane Proteins / Q000378:metabolism; D051381:Rats; D011916:Rats, Inbred F344; D033581:Stem Cell Transplantation; D013997:Time Factors
C512923:Dlk1 protein, rat; D036341:Intercellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D018819:Dipeptidyl Peptidase 4