Flow-cytometric determination of neutral lipids
Scope of the method
- Human health
- Translational - Applied Research
- In vitro - Ex vivo
- Human derived cells / tissues / organs
- neutral lipids
- in vitro
- stem cells
- lipid accumulation
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)
- Cell culture equipment;
- Biosafety cabinet.
- Internally validated
Pros, cons & Future potential
Measuring many samples can be time-consuming.
You can use also other cell types.
- Drug testing (anti-steatotic drugs) ;
- Assessing drug-induced liver steatosis (in vitro).
References, associated documents and other information
M. T. Donato et al., Chem. Biol. Interact. 181, 417–423 (2009)
R. M. Rodrigues et al., Stem Cells Dev. 23, 44–55 (2014)
Contact personJoost Boeckmans
OrganisationsVrije Universiteit Brussel (VUB)
Pharmaceutical and Pharmacological Sciences
In Vitro Toxicology and Dermato-Cosmetology