Jan 04, 2023
Version 1
0.1% BSA media prep and flow V.1
- 1Max Delbrück Center for Molecular Medicine
Protocol Citation: Olya Oppenheim 2023. 0.1% BSA media prep and flow. protocols.io https://dx.doi.org/10.17504/protocols.io.x54v9d7zpg3e/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: January 02, 2023
Last Modified: January 04, 2023
Protocol Integer ID: 74646
Keywords: bmp pathway, flow experiments with the ibidi perfusion system, ibidi perfusion system, fetal bovine serum, onset of flow, replacing egm2, egm2, flow, flow experiment
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.
BSA 0.1% media prep
5m
50 mL EGM2 without FBS
667 µL BSA 7.5% solution
filtrate through 0.22um yellow filter.
Add to slides 4h before starting flow.
Optional
adding BMP9/10 to flow experiment
in order to obtain a final 0.5 ng/ml , add 1.4ul BMP9/10 of 10 ug/ml initial concentration per perfusion unit (volume of 14ml)
add this to each perfusion unit during slide connection