Expressions of hepatobiliary organic anion transporters and bilirubin-conjugating enzyme in differentiating embryonic stem cells.
Mouse embryonic stem (ES) cells are clonal cell lines derived from the inner cell mass of developing blastocysts and have multi-lineage differentiation ability. We previously reported that ES cells can be made to differentiate into hepatocytes possessing high metabolic activities by transfection of hepatocyte nuclear factor-3beta (HNF-3beta). In the present study, we investigated the expression of hepatobiliary organic anion transporters and bilirubin uridine diphosphate glucuronosyltransferase (ugt1a1) in undifferentiated and differentiating HNF-3beta-transfected ES (HNF-3beta-ES) cells. The expression of organic anion transporting polypeptide 1 (oatp1), multidrug resistance-associated protein 1 (mrp1), mrp2, mrp3, and ugt1a1 was not seen in the undifferentiated HNF-3beta-ES cells by RT-PCR, whereas all were expressed in differentiating HNF-3beta-ES cells. Protein expression for oatp1, mrp1, mrp2, mrp3, and ugt1a1 was also observed in the differentiating HNF-3beta-ES cells by Western blotting. An immunofluorescence examination revealed that oatp1 was co-located with desmoplakin, a marker for the basolateral (sinusoidal) membrane, and mrp2 was co-localized with CD26, a marker for the apical (canalicular) membrane, though they were both expressed throughout most of the cell membranes.
Tanaka
Yuji
Y
Third Department of Internal Medicine, Mie University School of Medicine, 2-174 Edobashi, Tsu city 514-8507, Japan.
Yoshikawa
Masahide
M
Kobayashi
Yoshinao
Y
Kuroda
Makoto
M
Kaito
Masahiko
M
Shiroi
Akira
A
Yamao
Jun-ichi
J
Fukui
Hiroshi
H
Ishizaka
Shigeaki
S
Adachi
Yukihiko
Y
D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D014453:Glucuronosyltransferase / Q000096:biosynthesis* / Q000235:genetics; D051542:Hepatocyte Nuclear Factor 3-beta; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism*; D051379:Mice / Q000196:embryology / Q000235:genetics; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000096:biosynthesis* / Q000235:genetics; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D027361:Organic Anion Transporters / Q000096:biosynthesis* / Q000235:genetics; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*; D014157:Transcription Factors; D014162:Transfection / Q000379:methods; D000099148:UGT1A1 Enzyme
D004268:DNA-Binding Proteins; D014453:Glucuronosyltransferase; D051542:Hepatocyte Nuclear Factor 3-beta; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D009687:Nuclear Proteins; D027361:Organic Anion Transporters; D014157:Transcription Factors; D000099148:UGT1A1 Enzyme; C496503:Foxa2 protein, mouse