Isolation, characterization and culture of Thy1-positive cells from fetal rat livers.
OBJECTIVE
To investigate whether Thy1 recognizes oval cells in the fetal liver and to characterize the cultured Thy1- selected cells from E14 rat livers.


METHODS
Thy1 populations were analyzed by fluorescence activated cell sorter analysis. Thy1 positive cells were isolated using magnetic beads. Hepatic markers were detected by Western blotting, immunocytochemistry and RT-PCR.


RESULTS
The percentage of Thy1-positive cells decreased during early development of fetal rat liver (E13-E16). E14 fetal livers contained 7.8% Thy1 positive cells, of which 61% were positive for alpha-fetoprotein (AFP) and 25% expressed albumin. The Thy1+ population expressed oval cell markers c-Kit and CXCR4, liver enriched-transcription factors HNF1alpha and HNF6, hepatocytic markers albumin, AFP and cytokeratin 18, and biliary marker cytokeratin 19. Thy1- selected cells formed only mesenchymal colonies when plated on collagen and in serum-containing media. Thy1 selected cells were able to form hepatic colonies positive for HNF1alpha, HNF6, albumin, AFP, cytokeratin 18, cytokeratin 19 and glycogen, when grown on STO feeder layers in serum free-media.


CONCLUSIONS
Oval cells positive for Thy1 are present in early liver embryonic stages.
Isabel
Zvibel
Z
Liver Unit, Gastroenterology Institute, Tel Aviv Sourasky Medical Center, Israel. isab@tasmc.health.gov.il
Miri
Bronstein
B
Einav
Hubel
H
Ella
Bar-Lev
BL
Zamir
Halpern
H
Ran
Oren
O
D000418:Albumins / Q000032:analysis; D000818:Animals; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D005260:Female; D051538:Hepatocyte Nuclear Factor 1-alpha / Q000032:analysis; D051559:Hepatocyte Nuclear Factor 6 / Q000032:analysis; D022781:Hepatocytes / Q000737:chemistry / Q000166:cytology* / Q000276:immunology*; D007150:Immunohistochemistry; D007633:Keratins / Q000032:analysis; D008099:Liver / Q000737:chemistry / Q000166:cytology / Q000196:embryology* / Q000276:immunology*; D020930:MAP Kinase Kinase Kinases / Q000032:analysis; D011247:Pregnancy; D051381:Rats; D011916:Rats, Inbred F344; D018121:Receptors, Cytokine / Q000032:analysis; D053677:Receptors, Oncostatin M; D018800:Thy-1 Antigens / Q000032:analysis*; D000509:alpha-Fetoproteins / Q000032:analysis
D000418:Albumins; D051538:Hepatocyte Nuclear Factor 1-alpha; D051559:Hepatocyte Nuclear Factor 6; C496489:Hnf1a protein, rat; C496607:Onecut1 protein, rat; D018121:Receptors, Cytokine; D053677:Receptors, Oncostatin M; D018800:Thy-1 Antigens; D000509:alpha-Fetoproteins; D007633:Keratins; D020930:MAP Kinase Kinase Kinases; C090757:mitogen-activated protein kinase kinase kinase 12