Derivation and Characterization of Hepatic Progenitor Cells from Human Embryonic Stem Cells

Abstract

The derivation of hepatic progenitor cells from human embryonic stem (hES) cells is of value both in the study of early human liver organogenesis and in the creation of an unlimited source of donor cells for hepatocyte transplantation therapy. Here, we report for the first time the generation of hepatic progenitor cells derived from hES cells. Hepatic endoderm cells were generated by activating FGF and BMP pathways and were then purified by fluorescence activated cell sorting using a newly identified surface marker, N-cadherin. After co-culture with STO feeder cells, these purified hepatic endoderm cells yielded hepatic progenitor colonies, which possessed the proliferation potential to be cultured for an extended period of more than 100 days. With extensive expansion, they co-expressed the hepatic marker AFP and the biliary lineage marker KRT7 and maintained bipotential differentiation capacity. They were able to differentiate into hepatocyte-like cells, which expressed ALB and AAT, and into cholangiocyte-like cells, which formed duct-like cyst structures, expressed KRT19 and KRT7, and acquired epithelial polarity. In conclusion, this is the first report of the generation of proliferative and bipotential hepatic progenitor cells from hES cells. These hES cell–derived hepatic progenitor cells could be effectively used as an in vitro model for studying the mechanisms of hepatic stem/progenitor cell origin, self-renewal and differentiation.

Promotion of the efficient metabolic maturation of human pluripotent stem cell-derived hepatocytes by correcting specification defects

“…After the induction of hESCs through definitive endoderm cells [8] to ventral foregut cells ( Figure 1A and Supplementary information, Figures S2 and S3), the cells are recruited to the hepatic lineage. Because both BMP and FGF signals were included in our previous protocol [5], we postulated that an additional signal is required for hepatic PROX1 and HNF6 expression.…”

Take a deep breath and digest the material: organoids and biomaterials of the respiratory and digestive systems

“…Deriving biliary organoids will allow for an in vitro human model of the drug secretion system and biliary gland diseases. [102][103][104][105] For new and current drugs, it is crucial to understand the process of elimination through the biliary gland, which could help facilitate drug development and design, [106,107] and the biliary organoids could provide a system to model this process. In addition to drug secretion, biliary gland organoids derived from iPSC lines can model genetic diseases such as cystic fibrosis (CF) where the cystic fibrosis transmembrane conductance regulator (CFTR) is impaired and disrupts fluid secretion within the glands.…”

“…[86,109] These cells have been derived from hPSC and liver progenitor cell lines. [102][103][104][105]110] The liver and biliary gland are derived from the same progenitor population during development, called hepatoblasts. [86,109,111] This population has been derived from hPSCs and has the capability of becoming either hepatocytes or cholangiocytes depending on the growth factors added to the culture.…”

“…The hPSC-derived cholangiocytes can be cultured in a three-dimensional environment that consists of Matrigel and Collagen type I. [102][103][104][105] In the three-dimensional environment, the hPSC-derived cholangiocytes had proper cellular organization forming an apical-basal polarity with barrier function, similar to native cholangiocytes. [102][103][104][105]110] When hPSC-derived cholangiocytes were co-cultured with Op9 stromal cell to stimulate Notch signaling, ductal, and tubular structures formed similar to native bile ducts.…”

“…[102] Cholangiocytes derived without stromal cells did form the apical-basal polarity, but resembled a cyst-like structure rather than the native ductal structures. [103][104][105] To model the secretion system, the cell barrier with the MDR1 transporter was tested as a function of the biliary gland. [102][103][104][105]110] Thus, an intact cell barrier is crucial to the biliary gland function.…”