A small molecule that directs differentiation of human ESCs into the pancreatic lineage

Abstract

Stepwise differentiation from embryonic stem cells (ESCs) to functional insulin-secreting beta cells will identify key steps in beta-cell development and may yet prove useful for transplantation therapy for diabetics. An essential step in this schema is the generation of pancreatic progenitors--cells that express Pdx1 and produce all the cell types of the pancreas. High-content chemical screening identified a small molecule, (-)-indolactam V, that induces differentiation of a substantial number of Pdx1-expressing cells from human ESCs. The Pdx1-expressing cells express other pancreatic markers and contribute to endocrine, exocrine and duct cells, in vitro and in vivo. Further analyses showed that (-)-indolactam V works specifically at one stage of pancreatic development, inducing pancreatic progenitors from definitive endoderm. This study describes a chemical screening platform to investigate human ESC differentiation and demonstrates the generation of a cell population that is a key milepost on the path to making beta cells.

CD24: A Novel Surface Marker for PDX1-Positive Pancreatic Progenitors Derived from Human Embryonic Stem Cells

“…Significant progresses have been made in directing human ESCs to differentiate into pancreatic progenitors and insulin-producing beta cells by stepwise induction [2][3][4][5][6][7][8]. However, although pancreatic progenitors can be efficiently generated from human ESCs, efficient in vitro differentiation into mature functional beta cells that are able to reverse diabetes in animal models has not been achieved.…”

Embryonic stem cell application in drug discovery

“…Cell-based in vitro assays with high human relevance are urgently needed for pre-clinical activities. Previous studies have suggested that ES cells could serve as a screening platform to identify low-molecular-weight compounds that affect endogenous stem cell populations and repair damaged tissue [4,8,10,11,28,29] , and a set of screening protocols is available (ie, the primary screen and the secondary assay). In addition, several cell lines, such as D3 mES cells and H1, H7, and H9 hES cells, are available for drug screening [12,30,31] .…”

“…As a result, several steroids that promoted hES cell differentiation were identified, and the antihypertensive drug pinacidil was shown to affect hES cell survival. Phenotypic screens can be advantageous because they can be carried out in cells by examining multiple markers and functional changes (for example, cell morphology and behavior) using automated high-content imaging technologies in a high-throughput manner [1,4,36,37] .…”

“…An outstanding landmark in the past decade has been the development between stem cell biology and chemistry [1][2][3][4][5][6] . The combination of the two fields will likely provide an impetus for revealing intricate molecular interactions and functions under complex cellular differentiation, which offers systems for drug discovery and sources for potential cell therapy [7][8][9][10] .…”

“…This makes them potentially valuable for pharmaceutical development and safety assessment. Indeed, small molecules can be screened using hES cell models to find new chemical entities that modulate the fate of adult stem cells, or they could be used directly in cell therapies [4,8,10,11] .…”