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Revolutionizing Liver Disease Treatment with iPSC-Derived Liver Organoids

Did you know scientists at the University of Tokyo and Yokohama City University have developed an animal-derived product free protocol for mass production of mini-livers, revolutionizing clinical trials and treatment for liver diseases?

Human induced pluripotent stem cells (iPSCs) have emerged as a promising tool in regenerative medicine, with successful transplantation into patients over the past decade. The production of iPSC-derived liver buds (iPSC-LBs) has presented the exciting potential of new avenues for the treatment of liver diseases such as urea cycle disorders and liver cirrhosis. However, taking iPSC-LBs from the laboratory to the clinic has required the challenging development of a protocol involving animal-free products to meet the regulatory standards for therapeutic applications.

Ajinomoto has collaborated with Professor Taniguchi and his research group at The University of Tokyo’s Department of Regenerative Medicine to develop StemFit® for Differentiation (stocked by AMSBIO), a chemically-defined, animal-origin free media (CD-AOF) for iPSC differentiation. Using this innovative product Prof. Taniguchi and his team have successfully developed an entirely animal-free protocol for differentiating iPSCs into three cell types to generate iPSC-LBs. Building on this breakthrough, Dr. Satoshi Okamoto and his team at Yokohama City University Graduate School of Medicine’s Department of Regenerative Medicine have effectively utilised this protocol to achieve larger-scale stable production of iPSC-LBs for clinical applications.

Figure 1. Schematic representation of the generation of liver buds and the media. Image from Sekine, K., Ogawa, S., Tsuzuki, S., Kobayashi, T., Ikeda, K., Nakanishi, N., Takeuchi, K., Kanai, E., Otake, Y., Okamoto, S., Kobayashi, T., Takebe, T., & Taniguchi, H. (2020). Generation of human induced pluripotent stem cell-derived liver buds with chemically defined and animal origin-free media. Scientific Reports, 10(1). https://doi.org/10.1038/s41598-020-73908-1

“…By using StemFit® AK03N (StemFit® Basic03) for iPSC cell culture and StemFit® for Differentiation for differentiation, we were able to adopt animal-origin free reagents, which made the culture protocol very stable.” – Dr. Satoshi Okamoto, Yokohama City University graduate School of Medicine

Prof. Taniguchi’s and Dr. Okamoto’s groups maintained their iPSC cell culture in StemFit® Basic03, on plates coated with iMatrix-511 recombinant laminin, both available from AMSBIO. The researchers replaced conventional differentiation media which used animal-derived products with a media supplemented with StemFit® for Differentiation and StemFit Purotein Human recombinant Activin A. The evaluation of the new CD-AOF medium protocol demonstrated comparable efficiency to the conventional medium in differentiating iPSCs into definitive endoderm (DE), hepatic endoderm (HE), and immature hepatocyte (IH) stages, as well as the differentiation of IHs to mature hepatocytes (MHs). Additionally, the newly developed protocol led to improved albumin secretion and higher expression of several mature hepatic markers in MHs compared to the conventional protocol.

Figure 2. Lipid incorporation study in all iPSC-LBs by using DiI-Ac-LDL (Acetylated Low Density Lipoprotein labeled with 1,1’-dioctadecyl-3,3,3’,3’- tetramethylindo-carbocyanine perchlorate). Scale bar, 50 µm. Figure adapted from Sekine, K., Ogawa, S., Tsuzuki, S., Kobayashi, T., Ikeda, K., Nakanishi, N., Takeuchi, K., Kanai, E., Otake, Y., Okamoto, S., Kobayashi, T., Takebe, T., & Taniguchi, H. (2020). Generation of human induced pluripotent stem cell-derived liver buds with chemically defined and animal origin-free media. Scientific Reports, 10(1). https://doi.org/10.1038/s41598-020-73908-1

“StemFit® For Differentiation allows for stable differentiation induction and is very easy to use.” – Dr. Satoshi Okamoto, Yokohama City University graduate School of Medicine

iPSC-LBs generated using the CD-AOF medium exhibited consistent circularity and minimal size variation, showcasing functional maturation during the comprehensive functional analysis. Comparative gene expression analysis of the iPSC-LBs to liver tissue and primary human hepatocytes showed that the CD-AOF samples did not display significantly inferior gene expression levels, except for AAT and TAT. Notably, the higher albumin secretion produced by CD-AOF LBs observed in vitro also led to increased albumin expression in vivo when iPSC-LBs were transplanted into mice. These findings underscore the potential of iPSC-LBs and the effectiveness of the CD-AOF approach.

“We believe that the reason why we have been able to achieve stable culture even in scale-up is because we have been using this supplement to build a differentiation induction system from the small-scale stage, and also because StemFit® for Differentiation is a chemically-defined supplement.” – Dr. Satoshi Okamoto, Yokohama City University graduate School of Medicine

In an interview with Ajinomoto, Dr. Okamoto describes how using the protocol he helped develop with Prof. Taniguchi, he and his team have continued to refine and expand the large-scale production of CD-AOF iPSC-LBs in compliance with the Pharmaceutical and Medical Devices Agency regulations. Moving forward, Dr. Sakamoto and his fellow researchers are working on enhancing the stability of iPSC-LBs during transport and storage. They are also exploring ways to improve iPSC-LBs by modifying the constituent cells. Their goal with this research is to create iPSC-LBs with enhanced functions, enabling their application in a broader range of liver diseases. AMSBIO is proud to stock the complete StemFit® range. Available in both research and clinical-grade formulations, StemFit® provides exceptional colony-forming efficiency while minimizing media volume usage making it the most cost-effective choice compared to other leading competitors in the market. Head to our page and discover how StemFit® can streamline your research.

iPSCs – Induced pluripotent stem cells
iPSC-LBs – Induced pluripotent stem cell-derived liver buds
CD-AOF – Chemically-defined animal origin-free
DE – Definitive endoderm
HE – Hepatic endoderm
IH – Immature hepatocyte
MHs – Mature hepatocyte
AAT – A gene that makes the protein Alpha-1 antitrypsin (AAT)
TAT – A gene that makes the liver enzyme tyrosine aminotransferase (TAT)

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We are a global provider of human and animal biospecimens: including frozen & FFPE tissue, DNA, RNA, total proteins, blood products and primary cells.

We are a global provider of human and animal biospecimens: including frozen & FFPE tissue, DNA, RNA, total proteins, blood products and primary cells.

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We are a global provider of human and animal biospecimens: including frozen & FFPE tissue, DNA, RNA, total proteins, blood products and primary cells.

We are a global provider of human and animal biospecimens: including frozen & FFPE tissue, DNA, RNA, total proteins, blood products and primary cells.

We are a global provider of human and animal biospecimens: including frozen & FFPE tissue, DNA, RNA, total proteins, blood products and primary cells.

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The CELLBANKER® series of cryopreservation media allows for the stable long-term storage of any cell type, including sensitive cell lines.

The CELLBANKER® series of cryopreservation media allows for the stable long-term storage of any cell type, including sensitive cell lines.

The CELLBANKER® series of cryopreservation media allows for the stable long-term storage of any cell type, including sensitive cell lines.

The CELLBANKER® series of cryopreservation media allows for the stable long-term storage of any cell type, including sensitive cell lines.

The CELLBANKER® series of cryopreservation media allows for the stable long-term storage of any cell type, including sensitive cell lines.

The CELLBANKER® series of cryopreservation media allows for the stable long-term storage of any cell type, including sensitive cell lines.

The CELLBANKER® series of cryopreservation media allows for the stable long-term storage of any cell type, including sensitive cell lines.

The CELLBANKER® series of cryopreservation media allows for the stable long-term storage of any cell type, including sensitive cell lines.

The CELLBANKER® series of cryopreservation media allows for the stable long-term storage of any cell type, including sensitive cell lines.

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AMSBIO offers one of the largest selection of active recombinant enzymes and enzyme activity assays for profiling, inhibitor screening assays, and drug discovery.

AMSBIO offers one of the largest selection of active recombinant enzymes and enzyme activity assays for profiling, inhibitor screening assays, and drug discovery.

AMSBIO offers one of the largest selection of active recombinant enzymes and enzyme activity assays for profiling, inhibitor screening assays, and drug discovery.

AMSBIO offers one of the largest selection of active recombinant enzymes and enzyme activity assays for profiling, inhibitor screening assays, and drug discovery.

AMSBIO offers one of the largest selection of active recombinant enzymes and enzyme activity assays for profiling, inhibitor screening assays, and drug discovery.

AMSBIO offers one of the largest selection of active recombinant enzymes and enzyme activity assays for profiling, inhibitor screening assays, and drug discovery.

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AMSBIO offers one of the largest selection of active recombinant enzymes and enzyme activity assays for profiling, inhibitor screening assays, and drug discovery.

AMSBIO offers one of the largest selection of active recombinant enzymes and enzyme activity assays for profiling, inhibitor screening assays, and drug discovery.

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We provide the most advanced method for gene and shRNA delivery with an extensive collection of viral particles as well as a range of custom services.

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Prepared from Arthrobacter luteus, Zymolyase has a highly effective lytic activity against yeast cell walls.

Prepared from Arthrobacter luteus, Zymolyase has a highly effective lytic activity against yeast cell walls.

Prepared from Arthrobacter luteus, Zymolyase has a highly effective lytic activity against yeast cell walls.

Prepared from Arthrobacter luteus, Zymolyase has a highly effective lytic activity against yeast cell walls.

Prepared from Arthrobacter luteus, Zymolyase has a highly effective lytic activity against yeast cell walls.

Cells in Space: Exploring Neurodegenerative Diseases in Microgravity

miRNA: Alleviating Anxiety in the Amygdala

Revolutionizing Liver Disease Treatment with iPSC-Derived Liver Organoids

Further reading

iMatrix Recombinant Laminin E8 fragments

StemFit Purotein® Recombinant Proteins

StemFit® Feeder-Free Stem Cell Culture Media

StemFit® For Differentiation

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