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A protocol for human pluripotent stem cell differentiation into endoderm related cells

 

Pluripotent cells like embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) are promising cell source for cell replacement therapies against diseases of endoderm derived organs, such as liver, lung and pancreas. However, most differentiation protocols does not provide sufficiently defined starting conditions, therefore, complicate the adaption of pluripotent cells. Diekmann et al. showed a detailed protocol started from a defined cell number of dispersed single cells of three different human ESC lines, and one human iPSC line. The article was published on Stem Cells and Development.

 

For the effective induction of definitive endoderm state, researchers activated ActivinA/Nodal signaling as well as inhibited GSK3 signaling for the first 24h, and then inhibited ActivinA/Nodal signaling. They found activation of ActivinA/Nodal signaling alone was not sufficient for the endoderm state commitment. The protocol provides a feeder-independent approach since feeder cells hindered the differentiation process. In addition, an inhibition of PI3K was not necessary. According to this protocol, ESCs and iPSCs were able to differentiated into PDX1-positive pan-pancreatic cells and NGN3-positive endocrine progenitors. This protocol provides a better strategy for efficient production of endoderm cells from feeder-free cultivated human pluripotent stem cells.

 

Reference:
Stem Cells Dev. 2015 Jan 15;24(2):190-204. 

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S1049 Y-27632 2HCl Y-27632 2HCl is a selective ROCK1 and ROCK2 inhibitor with a Ki of 140 nM and 300nM in a cell-free assay, exhibits >200-fold selectivity over other kinases, including PKC, cAMP-dependent protein kinase, MLCK and PAK.

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