Hepatocyte lineage cells derived from pluripotent stem cells

C - Chemistry – Metallurgy – 12 – N

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Details

C12N 5/071 (2010.01) C12N 5/00 (2006.01) C12N 5/0735 (2010.01) A61K 35/407 (2006.01)

Patent

CA 2407505

It has been discovered that when pluripotent stem cells are cultured in the presence of a hepatocyte differentiation agent, a population of cells is derived that has a remarkably high proportion of cells with phenotypic characteristics of liver cells. In one example, human embryonic stem cells are allowed to form embryoid bodies, and then combined with the differentiation agent n-butyrate, optionally supplemented with maturation factors. In another example, n-butyrate is added to human embryonic stem cells in feeder-free culture. Either way, a remarkably uniform cell population is obtained, which is predominated by cells with morphological features of hepatocytes, expressing surface markers characteristic of hepatocytes, and having enzymatic and biosynthetic activity important for liver function. Since stem cells readily proliferate in culture, this system provides an abundant source of cells of the hepatocyte lineage for a variety of applications, such as drug screening, and replenishing liver function in the context of clinical treatment.

On a découvert que lorsque des cellules souches pluripotentielles sont cultivées en présence d'un agent de différentiation d'hépatocyte, il est possible d'en dériver une population de cellules renfermant une proportion remarquablement élevée de cellules possédant des caractéristiques phénotypiques de cellules du foie. Selon un exemple, des cellules souches embryonnaires humaines peuvent former des corps embryonnaires, puis se combiner à l'agent de différentiation n-butyrate, éventuellement complété par des facteurs de maturation. Selon un autre exemple, n-butyrate est ajouté aux cellules souches embryonnaires humaines dans une culture exempte de cellules nourricières. Selon l'un ou l'autre exemple, on obtient une population cellulaire remarquablement uniforme dans laquelle prédominent des cellules présentant les caractéristiques morphologiques des hépatocytes, exprimant des caractéristiques de marqueurs de surface des hépatocytes, et possédant une activité enzymatique et biosynthétique importante pour une fonction du foie. Du fait que les cellules souches prolifèrent rapidement en culture, ce système fournit une source abondante de cellules de lignée d'hépatocytes pour une variété d'applications, telles que le criblage d'un médicament, et la reconstitution d'une fonction hépatique dans le contexte d'un traitement clinique.

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