Small Molecules Efficiently Direct Endodermal Differentiation of Mouse and Human Embryonic Stem Cells

Abstract

Summary An essential step for therapeutic and research applications of stem cells is the ability to differentiate them into specific cell types. Endodermal cell derivatives, including lung, liver and pancreas, are of interest for regenerative medicine, but efforts to produce these cells have been met with only modest success. In a screen of 4000 compounds, two cell permeable small molecules were indentified that direct differentiation of ESCs into the endodermal lineage. These compounds induce nearly 80% of ESCs to form definitive endoderm, a higher efficiency than that achieved with Activin A or Nodal, commonly used protein inducers of endoderm. The chemically induced endoderm expresses multiple endodermal markers, can participate in normal development when injected into the embryonic gut tube and can form pancreatic progenitors in vitro. The application of small molecules to differentiate mouse and human ESCs into endoderm, and pancreatic progenitors represents a step toward achieving a reproducible and efficient production of desired ES cell derivatives.

Connective tissue growth factor acts within both endothelial cells and β cells to promote proliferation of developing β cells

“…These protocols have been successful in generating insulin-producing cells; however, in general, the percentage of β cells in the cultures is relatively low and expansion of these cells has proven difficult (36). Recent studies have sought to identify small molecules or growth factors that are able to increase the yield of β cells (37). Our studies suggest that CTGF is an attractive candidate for inclusion in these directed differentiation protocols, as it is required for proper lineage allocation and embryonic β-cell proliferation and its overexpression in β cells was sufficient to increase endocrine proliferation and mass.…”

Differentiation of Stem Cells into Insulin-Producing Cells: Current Status and Challenges

“…Differentiation of ESCs into definitive endoderm can be achieved via treatment with activin-A and Wnt3a and confirmed by expression of endodermal markers, including SOX17, FOXA2, GATA4, CXCR4, and cerberus (D'Amour et al 2006;Kroon et al 2008). Nodal, CHIR99021, IDE1 and IDE2 are another molecules that induce development of the definitive endoderm (Borowiak et al 2009;Takenaga et al 2007). The next step after formation of definitive endoderm is induction of primitive gut tube.…”

Chemical approaches to studying stem cell biology

“…Another study used Sox17 promoter-driven dTomato as a reporter, and screened a collection of 4 000 compounds for small molecules that could induce definitive endoderm in the absence of activin A. Two structurally similar small molecules, IDE1 and IDE2, were found to induce endoderm differentiation in up to 80% of mESCs (or 50% of hESCs) in the absence of activin A [30]. Similar to activin A, both IDE1 and IDE2 induce SMAD2 phosphorylation in mESCs although their molecular targets remain unknown.…”