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Afamin/Wnt3a Condition Medium

Condition medium to help organoid formation

Why the Wnt signaling pathway is important

Wnt signaling is known to be involved in early development, maintenance and regeneration of stem cells, and in cancer formation. Wnt signaling has also been found to play an important role in the growth and maintenance of these processes. In particular, Wnt3a has been revealed to be an essential niche component for maintaining the proliferation of Lgr5-positive stem cells in intestinal epithelial cells and is used for the production of various digestive organoids such as the small intestine, large intestine, stomach, pancreas and liver. Although Wnt3a has been conventionally used for the culture of gut organoids, it is a fat-soluble protein, so it forms aggregates in serum-free medium and can not exert its activity sufficiently. In 2016, Mihara et al. found that high Wnt3a activity can be maintained by forming a complex with Wnt3a by Afamin, which is one of the components of serum. In addition, by using Afamin and Wnt3a complex for organoid culture, long-term culture of organoid becomes possible. This new medium will result in optimal success for your organoid experiments.

SEE AFAMIN/WNT3a PRODUCTS

Mechanism of Wnt3a Stabilization by Afamin Proteins

Afamin/Wnt3a CM increased Lgr5 Positive Stem Cells Expressed td Tomato

This data obtained from collaboration with Dr. Sato, Keio Univ

Images on the top panels show a bright field of human color organoids. Images on the bottom panels show fluorescent LGR5 positive stem cells that express td tomato regulated by Lgr5 promoter. Afamin/Wnt3a CM maintained LGR5 positive stem cell growth is seen at greater levels compared with cell growth in commercial recombinant Wnt3a (300ng/mL).

Recombinant Afamin/Wnt3a increased LGR5 Positive Stem Cells

An organoid strain expressing tdTomato regulated by Lgr5 promoter was cultured in the presence of Recombinant Afamin/Wnt3a and Recombinant Wnt3a of competitor A. It was confirmed that Recombinant Afamin/Wnt3a maintains LGR5-positive cells even at low concentrations.

What are the Differences Between Organoids and Spheroids?

In general, there are two major differences:

  1. Nature of the driving force for development: Whereas internal developmental processes drive organoid formation, spheroids develop primarily via cell-to-cell adhesion.
  2. Length of time 3D cultures can be maintained: Long term, in vitro expansion of cells in culture needs an immature stem cell population to replenish dying cells. Organoids are derived from, and maintain, a population of stem cells during in vitro culture, which guarantees their long term viability. This is achieved by optimizing culture growth conditions, such as providing a basement membrane matrix (i.e. Matrigel®), and adding a selection of agonists (e.g. Wnt and tyrosine kinase receptor) and inhibitors (e.g. bone morphogenetic protein/transforming growth factor-β).

Importantly, when organoids are passaged they retain the genetic features of the original organ over several generations. In contrast, long-term culture of tissue-derived spheroids is challenging, possibly due to inherent technical difficulties in extracting and maintaining viable cells.

Organoids and spheroids can be generated from a variety of healthy as well as diseased cell types and tissues, such as patient tumors. Tumor derived organoids and spheroids have been generated and extensively investigated for their use in drug discovery. However, there are some key differences in establishing the two from patient-derived tumors.

FGF-Max is our new product, shown to provide success with organoid cultures being acting as a universal FGF receptor agonist. Find out how this reagent works!

FGF-Max


Product Highlights

Product Code Product Name
J2-001 Afamin/Wnt3a CM
J2-002 Recombinant Afamin/Wnt3a
J2-003 FGF-Max(Recombinant human FGF-1/FGF-2 chimera)

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