Characterization of hepatic progenitors from human fetal liver during second trimester.
OBJECTIVE
To enrich hepatic progenitors using epithelial cell adhesion molecule (EpCAM) as a marker from human fetal liver and investigate the expression of human leukocyte antigen (HLA) and their markers associated with hepatic progenitor cells.


METHODS
EpCAM +ve cells were isolated using magnetic cell sorting (MACS) from human fetuses (n = 10) at 15-25 wk gestation. Expression of markers for hepatic progenitors such as albumin, alpha-fetoprotein (AFP), CD29 (integrin beta1), CD49f (integrin alpha6) and CD90 (Thy 1) was studied by using flow cytometry, immunocytochemistry and RT-PCR; HLA class I (A, B, C) and class II (DR) expression was studied by flow cytometry only.


RESULTS
FACS analysis indicated that EpCAM +ve cells were positive for CD29, CD49f, CD90, CD34, HLA class I, albumin and AFP but negative for HLA class II (DR) and CD45. RT PCR showed that EpCAM +ve cells expressed liver epithelial markers (CK18), biliary specific marker (CK19) and hepatic markers (albumin, AFP). On immunocytochemical staining, EpCAM +ve cells were shown positive signals for CK18 and albumin.


CONCLUSIONS
Our study suggests that these EpCAM +ve cells can be used as hepatic progenitors for cell transplantation with a minimum risk of alloreactivity and these cells may serve as a potential source for enrichment of hepatic progenitor.
Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad 500007, India.
Mekala-Subba
MS
Rao
Aleem-Ahmed
AA
Khan
Nyamath
N
Parveen
Mohammed-Aejaz
MA
Habeeb
Chittoor-Mohammed
CM
Habibullah
Gopal
G
Pande
D000418:Albumins / Q000378:metabolism; D000951:Antigens, Neoplasm / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000378:metabolism; D002478:Cells, Cultured; D000071858:Epithelial Cell Adhesion Molecule; D005260:Female; D053686:Fetal Stem Cells / Q000276:immunology / Q000378:metabolism*; D005434:Flow Cytometry; D006680:HLA Antigens / Q000378:metabolism; D022781:Hepatocytes / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D018189:Immunomagnetic Separation; D039503:Integrin alpha6 / Q000378:metabolism; D019012:Integrin beta1 / Q000378:metabolism; D007633:Keratins / Q000378:metabolism; D008099:Liver / Q000196:embryology / Q000276:immunology / Q000378:metabolism*; D010641:Phenotype; D011247:Pregnancy; D011262:Pregnancy Trimester, Second; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018800:Thy-1 Antigens / Q000378:metabolism; D000509:alpha-Fetoproteins / Q000378:metabolism
C097353:AFP protein, human; D000418:Albumins; D000951:Antigens, Neoplasm; D015815:Cell Adhesion Molecules; C499379:EPCAM protein, human; D000071858:Epithelial Cell Adhesion Molecule; D006680:HLA Antigens; D039503:Integrin alpha6; D019012:Integrin beta1; D018800:Thy-1 Antigens; D000509:alpha-Fetoproteins; D007633:Keratins