0.1% BSA media prep and flow
Olya Oppenheim
Published: 2023-01-04 DOI: 10.17504/protocols.io.x54v9d7zpg3e/v1
Abstract
Fetal Bovine Serum contains traces of ligands of the BMP pathway which affect the flow experiments with the ibidi perfusion system.
replacing EGM2 with 2% with EGM2 with 0.1% BSA 4h prior to onset of flow (with same media) allows cells to respond better and collectively to flow.
Steps
BSA 0.1% media prep
1.
50mL EGM2 without FBS
667µL BSA 7.5% solution
filtrate through 0.22um yellow filter.
Add to slides 4h before starting flow.
Optional
2.
adding BMP9/10 to flow experiment
in order to obtain a final 0.5 , add 1.4ul BMP9/10 of 10 initial concentration per perfusion unit (volume of 14ml)
add this to each perfusion unit during slide connection
