Onto 100 mm dish (NIPPON Genetics), 0.3×106 MSC-UCs were plated and the culture medium was replaced with the same medium supplemented with 4 ATFs on the next day. The final concentration of each protein was 0.25 nM, and the same procedure was repeated twice a day for 5 days. On day 7, treated cells were harvested by using Accutase (Millipore), and subjected to analysis. No supplements for hepatocytic differentiation such as insulin or transferrin were used.
Protocol detail
MSC-UC Stimulation with 4 ATFs
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Corresponding Organization : National Center for Global Health and Medicine
Other organizations : Juntendo University, Hiroshima University, Tokyo Metropolitan Institute of Medical Science
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Variable analysis
independent variables
- Concentration of 4 ATFs (0.25 nM each)
- Cellular response to ATF treatment
- Cell type (MSC-UCs)
- Cell seeding density (0.3×10^6 cells per 100 mm dish)
- Culture medium (without insulin or transferrin supplements)
- Timing of ATF treatment (once daily for 5 days)
- Negative control: Untreated MSC-UCs
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