2. Adipogenesis 2
3. Insulin Signaling in Adipogenesis
4. Differentiation Grow cells until confluent
Place cells on differentiation medium for 4 days.
Place cells on post differentiation medium for 5 days.
Change medium every two days.
Lipid formation is visible around Day 4/5.
5. Oil Red O (ORO) Fix cells with paraformaldehyde solution
Expose cells to ORO solution for 15 min
Wash off excess ORO
Elute lipid adhered ORO with isopropanol.
Quantify amount of ORO present with spectrophotometer.
6. Oil Red O
7. Experiment 1. Each group will get 4 wells of 3T3L1 cells.
2. Cells will be seeded in growth media [ ref. Task sheet]
3. Students will change the growth media with differentiation media.
4. 4 different types of differentiation media will be added to each well. They will have different conc. of insulin. Hence each group will have 4 treatments.
5. after 2 days TA will change differentiation media to growth medium.
Next week, lipid formation will be measured by oil red O. [ ref. Task sheet]