FGF-Max (Recombinant human FGF-1/FGF-2 chimera)

■ What is FGF-Max?
■ Product Summary
■ Quality Assay results
　・Quantitative luciferase reporter assay
　・Human small intestinal organoid cultures
　・Establishment of mouse gastric organoids
　・Establishment of mouse pancreatic organoids

The family of fibroblast growth factors (FGFs) are composed of the growth factors involved in a variety of vital phenomena, including development, differentiation, proliferation, and morphogenesis. FGF-1～FGF-10 transmits signals into cells by binding to the FGF receptor (FGFR). Previous publications have suggested the FGF/FGFR signaling pathway is deeply involved in many processes from embryonic development to adult homeostasis.
Activation of FGF/FGFR signaling pathway is also known to be important in organoid cultures. However, proper selection of different FGFR ligands for each organ type is necessary while organoid culture, complicating experimental set up and execution. Therefore, using the universal FGFR ligand “FGF-1”, which shows highly affinity to all the type of FGFR, is expected to be a simple and convenient method for organoid culture. However, FGF-1 is known to be thermally unstable, and at 37°C, which cell-culture is normally being held, the bio-activity is lost within 6 hours, including in the presence of heparin. These factors results in commercially available FGF1 being unable to be used in culture studies.
To solve the problem, MBL provides FGF-Max, a universal FGFR ligand which the thermal stability is highly enhanced by chimerizing FGF-1 and FGF-2.


Ornitz DM, Itoh N. Wiley Interdiscip Rev Dev Biol. (2015)

Quantitative luciferase reporter assay

A quantitative luciferase reporter assay (SRE Reporter, Luciferase, HEK293 Cell Line: BPS Bioscience) was performed to assess the activity, and stability of FGF-Max.

● Comparing with FGF-2
FGF-Max and FGF-2 (commercially available products) were added to the cells at different concentration and cultured for 6 hours. The result shows that FGF-Max has the bio-activity in the same level with FGF-2.

● Thermal stability evaluation
After pre-incubation of FGF-Max and FGF-2 (commercially available product) at 37°C for 24, 48, and 72 hours, they were added to the cells for bio-activity assessment. The results suggested that FGF-Max retained higher bio-activity after pre-incubation compared to FGF-2 which also proves that FGF-Max has higher thermostability than FGF-2.

Human small intestinal organoid cultures

Human small intestinal organoids were cultured in the presence of FGF-Max and FGF-2 (commercially available product). It was suggested that proliferation of organoids is higher in the presence of FGF-Max than FGF-2.

● Differentiation of human small intestinal organoids
Paneth cell, endocrine cell, and goblet cell markers of human small intestinal organoids cultured in the presence of FGF-Max (4 ng/mL) were stained.
The presence of Paneth cells, and differentiated endocrine cells and goblet cells in organoids was confirmed.

Establishment of mouse gastric organoids

Murine gastric organoids were cultured in the presence of FGF-Max and FGF-10 (commercially available product). Similar to human small intestinal organoids, it was suggested that proliferation of organoids is higher in the presence of FGF-Max.

● ATP assay
ATP-assay was performed to assess the cell viability. As a result, it suggested that the cell viability in organoid is higher with the presence of FGF-Max.

Establishment of mouse pancreatic organoids

Murine pancreatic organoids were cultured in the presence of FGF-Max and FGF-10 (commercially available product). Similar to the above results, it was suggested that proliferation of organoids is higher in the presence of FGF-Max.

● ATP assay
ATP-assay was performed to assess the cell viability. As a result, it suggested that the cell viability of organoid is higher in the presence of FGF-Max.