Hepatic Differentiation of Mouse iPS Cells in Vitro

Abstract

Induced pluripotent stem (iPS) cells are pluripotent and are able to unlimitedly proliferate in vitro. This technical breakthrough in creating iPS cells from somatic cells has noteworthy implications for overcoming the immunological rejection and the ethical issues associated with the derivation of embryonic stem cells from embryos. In the current work, we present an efficient hepatic differentiation of mouse iPS cells in vitro. iPS cells were cultured free floating to induce the formation of embryoid bodies (EB) for 5 days. EB were transferred to a gelatin-coated plate and treated with 100 ng/ml activin A and 100 ng/ml basic fibroblast growth factor (bFGF) for 3 days to induce definitive endoderm. Cells were further cultured for 8 days with 100 ng/ml hepatocyte growth factor (HGF) to generate hepatocytes. Characterization was performed by RT-PCR assay. Functional analysis for albumin secretion and ammonia removal was also carried out. iPS cellderived hepatocyte-like cells (iPS-Heps) were obtained at the end of the differentiation program. Expression levels of a gestational hepatocyte gene and lineage-specific hepatic genes intensified in iPS-Heps. The production of albumin increased in a time-dependent manner. iPS-Heps were capable of metabolizing ammonia. We present here instant hepatic differentiation of mouse iPS cells using combined 3-day treatments of activin A and bFGF with subsequent 8-day HGF. Our study will be an important step to generate hepatocytes from human iPS cells as a new source for liver-targeted cell therapies.

Nuclear Translocation of β-Catenin during Mesenchymal Stem Cells Differentiation into Hepatocytes Is Associated with a Tumoral Phenotype

“…Hepatocytes differentiation has been achieved using different types of stem cells, MSC [17], embryonic stem cells [18] or induced pluripotent stem cells [19]. However in these studies the role of Wnt/β-catenin activation during hepatogenesis is unclear.…”

A preliminary study for constructing a bioartificial liver device with induced pluripotent stem cell-derived hepatocytes

“…Differentiation was carried out in four stages as described previously, with minor modification [22]. …”

Recent advances in 2D and 3D in vitro systems using primary hepatocytes, alternative hepatocyte sources and non-parenchymal liver cells and their use in investigating mechanisms of hepatotoxicity, cell signaling and ADME

“…The use of somatic cell sources has grown in the past few years, and iPS-derived hepatocytes have been generated from a variety of species including mouse (Okita et al 2007; Sancho-Bru et al 2011), human (Takahashi et al 2007b; Yu et al 2007; Song et al 2009) and pig (Aravalli et al 2012). Recent technological advances in iPS derivation have evolved for the development of footprint and feeder-free lines (Iwamuro et al 2010; Nagaoka et al 2010; Yu et al 2011) and the use of non-integrating episomal vector reprogramming of the patient T-cell population (Brown et al 2010; Mack et al 2011). These iPS cells can be expanded and directly differentiated into hepatocytes in vitro and are capable of many hepatic functions (1) albumin secretion, glycogen storage, drug metabolism, drug transportation and lipogenesis (Rashid et al 2010; Si-Tayeb et al 2010; Chen et al 2012).…”