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Development of an embryoid body-based screening strategy for assessing the hepatocyte differentiation potential of human embryonic stem cells following single-cell dissociation.

TitleDevelopment of an embryoid body-based screening strategy for assessing the hepatocyte differentiation potential of human embryonic stem cells following single-cell dissociation.
Publication TypeJournal Article
Year of Publication2013
AuthorsGreenhough S, Bradburn H, Gardner J, Hay DC
JournalCell Reprogram
Volume15
Issue1
Pagination9-14
Date Published2013 Feb
ISSN2152-4998
KeywordsAlbumins, Antigens, Differentiation, Cell Differentiation, Cell Line, Embryoid Bodies, Hepatocytes, Humans, Pluripotent Stem Cells
Abstract

We have devised an embryoid body-based screening method for the selection of human embryonic stem cell (hESC) lines capable of forming functional hepatocyte-like cells (HLCs) after single-cell dissociation. The screening method highlighted one cell line from a panel of five that produced albumin-positive cells during embryoid body (EB) formation. Cell lines that did not produce albumin-positive cells during EB formation were shown to respond less well to directed differentiation following single-cell replating. Additionally, the seeding density of the pluripotent populations prior to differentiation was shown to exert a significant effect on the hepatic function of the final population of cells. In summary, we have developed a simple procedure that facilitates the identification of human hESC lines that tolerate single-cell replating and are capable of differentiating to HLCs. Although the hepatic function of cells produced by this method is ∼10-fold lower than our current gold standard stem cell-derived models, we believe that these findings represent an incremental step toward producing HLCs at scale.

DOI10.1089/cell.2012.0049
Alternate JournalCell Reprogram
PubMed ID23379579