Development of an Embryoid Body-Based Screening Strategy for Assessing the Hepatocyte Differentiation Potential of Human Embryonic Stem Cells Following Single-Cell Dissociation

作者：Greenhough Sebastian; Bradburn Helen; Gardner John; Hay David C^*

来源：Cellular Reprogramming, 2013, 15(1): 9-14.

DOI：10.1089/cell.2012.0049

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 similar to 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.

• 出版日期2013-2