Flow-cytometric determination of neutral lipids

Using this method you can measure the (relative) lipid load in human skin-derived stem cells differentiated towards hepatic cells. This method could also be applied on other cell types (e.g. HepG2), since it is based on the following publication: "M. T. Donato et al., Chem. Biol. Interact. 181, 417–423 (2009)." Briefly: 1. Aspirate medium from the cell culture 2. Incubate 10’ with TrypLE (200 µL/well for 24- multiwell format) 3. Add 500 µL pre-warmed PBS (37°C) to every well and harvest the sample 4. Rinse with 500 µL PBS 5. Centrifigate according to the cell type 6. Resuspend in 1 mL PBS (containing BODIPY(TM) 1:2500 (see publication above)) on ice 7. 5’ before measuring + 1 µL Hoechst (+homogenize by pipetting) 8. Dilute 1:10 (PBS (4 °C)) before measuring to limit background signal 9. Measure signal (up to 100.000 events)