Novel population of hepatocytes derived via definitive endoderm (de-hep) from human blastocysts stem cells

Abstract

The present disclosure relates to a novel hepatocyte-like cell progenitor and / or a novel hepatocyte-like cell derived via definitive endoderm from human blastocyst-derived stem (hBS) cells, to a method for the preparation of such cells and to the potential use of such cells in, e.g., pharmaceutical drug discovery and development, toxicity testing, cell therapy and medical treatment. In particular is presented a definitive endoderm derived hepatocyte-like cell with important liver-expressed marker genes and important metabolizing enzymes, as well as drug transporters.

Description

FIELD OF THE INVENTION[0001]The present invention relates to a novel hepatoctye-like cell population derived via definitive endoderm from human blastocyst-derived stem (hBS) cells, to a method for the preparation of such cells and to the potential use of such cells in e.g. pharmaceutical drug discovery and development, toxicity testing, cell therapy and medical treatment. The cells are denoted DE-hep cells herein.[0002]Furthermore, the DE-Hep cells exhibits properties of functional human hepatocytes which make them attractive for use in in vitro studies of hepatogenesis such as early liver development processes or liver-regenerative disorders or malformations.BACKGROUND OF THE INVENTION[0003]Pluripotent human stem cells are expected to revolutionize the accessibility to a variety of human cell types. The possibility to propagate pluripotent human blastocyst-derived stem (hBS) cells and subsequently differentiate them into the desired target cell types will provide a stable and virtu...

AI Technical Summary

Benefits of technology

[0070]As used herein, the term “Alcohol dehydrogenase 1” is intended to mean a type of dehydrogenase enzyme that facilitate the interconversion between alcohols and aldehydes or ketones with the reduction of NAD+ to NADH. Alcohol dehydrogenase 1 serves to break down alcohols which could otherwise be toxic.

[0090]As used herein the term “MDR” is intended to mean multi-drug resistance transporter. MDR 1 and 3 are members of the ATP-binding cassette (ABC) family of transporters and both are drug efflux transporters. MDR 1 is important in regulating the traffic of drugs, peptides and xenobiotics into the body and in protecting the body against xenobiotic insults and drug toxicity, while MDR 3 is essential for phospholipid secretion into bile.

[0174]Characteristic vi) relates to the percentage of cells in the cell population comprising DE-Hep cells that exhibit protein and / or gene expression of the drug transporter MDR1 in the cell population according to the invention. MDR1 stands for multi-drug resistance transporter 1 and is a member of the ATP-binding cassette (ABC) family of transporters. MDR 1 is important in regulating the traffic of drugs, peptides and xenobiotics into the body and in protecting the body against xenobiotic insults and drug toxicity. MDR 1 is in vivo situated on the apical side of hepatocytes towards the bile canaliculus. In one embodiment of the present invention at least 5% such as, e.g., at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, or at least 80% at least 90% or at least 95% of the cells in the cell population comprising DE-Hep cells exhibit protein and / or gene expression of MDR1.

Problems solved by technology

Liver failure and end-stage liver diseases are responsible for a huge amount of deaths around the world and is a major burden on the health care system.

However, the efficacy of this procedure is limited and connected to many complications such as infection or rejection.

Liver transplantation also suffers from shortage of available donor organs and the treated patients will very often be referred to lifelong immunosuppression.

Unfortunately, the complexity and function of the liver is not mirrored by any cell type available today.

The availability of primary human liver cells is very limited and the cells are also known to rapidly loose their normal phenotype and functional properties (i.e. within 24 hours) when used for in vitro applications.

Thus, many tests are still performed using animal material, even though liver metabolism is known to be species specific and thereby generating difficulties in predicting liver metabolism and toxicity in another species than the one tested.

A complication in the study of endoderm is that mammals possess extraembryonic endoderm.

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