Cell Expansion During Directed Differentiation of Stem Cells Toward the Hepatic Lineage.
The differentiation of human pluripotent stem cells toward the hepatocyte lineage can potentially provide an unlimited source of functional hepatocytes for transplantation and extracorporeal bioartificial liver applications. It is anticipated that the quantities of cells needed for these applications will be in the order of 109-1010cells, because of the size of the liver. An ideal differentiation protocol would be to enable directed differentiation to the hepatocyte lineage with simultaneous cell expansion. We introduced a cell expansion stage after the commitment of human embryonic stem cells to the endodermal lineage, to allow for at least an eightfold increase in cell number, with continuation of cell maturation toward the hepatocyte lineage. The progressive changes in the transcriptome were measured by expression array, and the expression dynamics of certain lineage markers was measured by mass cytometry during the differentiation and expansion process. The findings revealed that while cells were expanding they were also capable of progressing in their differentiation toward the hepatocyte lineage. In addition, our transcriptome, protein and functional studies, including albumin secretion, drug-induced CYP450 expression and urea production, all indicated that the hepatocyte-like cells obtained with or without cell expansion are very similar. This method of simultaneous cell expansion and hepatocyte differentiation should facilitate obtaining large quantities of cells for liver cell applications.
1 Department of Chemical Engineering and Materials Science, University of Minnesota , Minneapolis, Minnesota.|2 Stem Cell Institute, University of Minnesota , Minneapolis, Minnesota.
Ravali
R
Raju
1 Department of Chemical Engineering and Materials Science, University of Minnesota , Minneapolis, Minnesota.|2 Stem Cell Institute, University of Minnesota , Minneapolis, Minnesota.|3 Department of Biomedical Engineering, University of Minnesota , Minneapolis, Minnesota.
David
D
Chau
1 Department of Chemical Engineering and Materials Science, University of Minnesota , Minneapolis, Minnesota.|2 Stem Cell Institute, University of Minnesota , Minneapolis, Minnesota.
Dong Seong
DS
Cho
1 Department of Chemical Engineering and Materials Science, University of Minnesota , Minneapolis, Minnesota.|2 Stem Cell Institute, University of Minnesota , Minneapolis, Minnesota.
Yonsil
Y
Park
4 Department of Development and Regeneration, Stem Cell Institute Leuven , KU Leuven, Leuven, Belgium .
Catherine M
CM
Verfaillie
1 Department of Chemical Engineering and Materials Science, University of Minnesota , Minneapolis, Minnesota.|2 Stem Cell Institute, University of Minnesota , Minneapolis, Minnesota.
Wei-Shou
WS
Hu
D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D019070:Cell Lineage / Q000235:genetics; D049109:Cell Proliferation / Q000235:genetics; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D006801:Humans; D008099:Liver / Q000166:cytology*; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
D015415:Biomarkers; D012333:RNA, Messenger
differentiation; embryonic stem cells; gene expression; genomics; hepatic