Differentiation of mouse and human embryonic stem cells into hepatic lineages

Abstract

We recently reported a novel method to induce embryonic stem (ES) cells differentiate into an endodermal fate, especially pancreatic, using a supporting cell line. Here we describe the modified culture condition with the addition and withdrawal of secreted growth factors could induce ES cells to selectively differentiate into a hepatic fate efficiently. The signaling of BMP and FGF that have been implicated in hepatic differentiation during normal embryonic development are shown to play pivotal roles in generating hepatic cells from the definitive endoderm derived from ES cells. Moreover, the expression of AFP, Albumin or a biliary molecular marker appeared sequentially thus suggested the differentiation of ES cells recapitulated normal developmental processes of liver. The ES cell‐derived differentiated cells showed evidence of glycogen storage, secreted Albumin, exhibited drug metabolism activities and expressed a set of cytochrome or drug conjugate enzymes, drug transporters specifically expressed in mature hepatocytes. With the same procedure, human ES cells also gave rise to cells with mature hepatocytes’ characteristics. In conclusion, this novel procedure for hepatic differentiation will be useful for elucidation of molecular mechanisms of hepatic fate decision at gut regionalization, and could represent an attractive approach for a surrogate cell source for pharmaceutical studies such as toxicology.

Directed hepatic differentiation from embryonic stem cells

“…Then Kume successfully developed an approach of inducing both mouse and human embryonic stem cells into mature hepatocytes by adding activin, bFGF, HGF, OSM, and Dex at different time points based on the original method of inducing embryonic stem cells into pancreatic cells (Agarwal et al, 2008). Lanza optimized Kume's method to differentiate mouse and human ESCs by adjusting the time points and the sequence of induction factors, and the efficiency of producing mature hepatocytes increased to 70%-80% (Shiraki et al, 2008). Basma et al were able to obtain functional hepatocytes from human ESCs using a similar protocol (Basma et al, 2009 this method with additional factors such as BMP4, FGF10 and retinoic acid, and they produced human hepatocytes exhibiting hepatic functions including glycogen storage, cytochrome activity and low-density lipoprotein uptake (Touboul et al, 2010).…”

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“…The consequences of these assays were (1) not reliable, (2) did not assess what they were assumed to measure, (3) extracted from cell types not relevant to the human system, (4) derived from the wrong conditions for measuring toxicities that occur in vivo at different conditions, and (5) in many cases, were misinterpreted. In vitro assays carry many limitations to measure ''epigenetic toxicants'' or, as some refer to them as ''non genotoxicants''.…”

“…Submit Manuscript | http://medcraveonline.com on these particular cell types can aid scientists in discovering treatment choices for different issue, not necessarily just using stem cells themselves to treat the disorder [2].…”

BMPS and Liver: More Questions than Answers

“…Furthermore, BMP4 has been reported to be necessary to induce definitive endoderm from mouse and human embryonic stem cells in vitro [80]. BMP2 appears to have a similar effect to that of BMP4 [81,82], whereas BMP7 has been shown to potentiate mesoderm differentiation and inhibit definitive endoderm differentiation [79,83]. Oval cells are adult liver stem cells that resemble embryonic hepatoblasts as they express both hepatocyte and bile duct markers.…”