FGF-Max: Quality Assay Results

Quantitative luciferase reporter assay

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

1. Comparing with FGF-2

FGF-Max and FGF-2 were added to the cells at different concentrations and cultured for 6 hours. The results show that FGF-Max has the same level of bio-activity as FGF-2.

2. Thermal stability evaluation

After pre-incubation of FGF-Max and FGF-2 at 37°C for 24 hours, 48 hours, 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.

3. Human small intestinal organoid cultures

Human small intestinal organoids were cultured in the presence of FGF-Max and FGF-2. The proliferation of organoids was higher in the presence of FGF-Max than in FGF-2.

4. Differentiation of human small intestinal organoids

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

Establishment of mouse gastric organoids

Murine gastric organoids were cultured in the presence of FGF-Max and FGF-10. Similar to human small intestinal organoids, the proliferation of organoids was shown to be higher in the presence of FGF-Max.

1. ATP assay

ATP-assay results suggested that the cell viability in organoids 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. Similar to the above results, it was suggested that the proliferation of organoids was higher in the presence of FGF-Max.