Methods Mol Biol. 2019;1994:101-115. doi: 10.1007/978-1-4939-9477-9_9.

Differentiation of hiPS Cells into Definitive Endoderm for High-Throughput Screening.

Bluhmki T¹, Traub S², Schruf E³, Garnett J³, Gantner F⁴, Bischoff D¹, Heilker R⁵.

Abstract

In drug discovery, there is an increasing demand for more physiological in vitro models that recapitulate the disease situation in patients. Human induced pluripotent stem (hiPS) cell-derived model cells could serve this purpose. To date, several directed differentiation approaches have been described to generate definitive endoderm (DE) from hiPS cells, but protocols suitable for drug development and high-throughput screening (HTS) have not been reported yet. In this work, a large-scale expansion of hiPS cells for high-throughput adaption is presented and an optimized stepwise differentiation of hiPS cells into DE cells is described. The produced DE cells were demonstrated to express classical DE markers on the gene expression and protein level. The here described DE cells are multipotent progenitors and act as starting points for a broad spectrum of endodermal model cells in HTS and other areas of drug discovery.