Exosome Isolation From Cell Culture Media Protocol

This chapter describes a detailed protocol for isolating exosomes from cell culture conditioned medium using ultracentrifugation and their characterization based upon size number and protein expression by several complementary methods such as transmission electron microscopy nanoparticle tracking analysis western blotting and flow cytometry.

Exosome isolation from cell culture media protocol.

Transfer the supernatant containing the cell free culture media to a new tube without disturbing the pellet. Simplifying methods for exosome enrichment and analysis. Exosomes 50 150 nm are extracellular vesicular structures secreted by cells in culture and found in body fluids 1. The yield of each reaction can yield pure exosome suspended in 50 200 µl pbs.

Each reaction can process 2 4 ml cell culture medium. Norgen s cell culture media exosome purification and rna isolation mini kit constitutes an all in one system for the purification of exosomes and the subsequent isolation of exosomal rna from different cell culture media sample volumes ranging from 5 ml to 10 ml. Repeated ultracentrifugation steps can reduce the quality of exosome preparations leading to lower exosome yield. Direct isolation of exosomes from cell culture.

Centrifuge the cell media at 2000 g. Technical report optimized exosome isolation protocol for cell culture supernatant and human plasma richard j. P100 kit is for exosome purification isolation at high yield and purity from cell culture media. 2school of medicine university of queensland.

Transfer the required volume of cell free culture media to a new tube and add 0 5 volumes of the total exosome isolation from. Lobb1 2 melanie becker1 shu wen wen1 christina s. Uncompromising ev isolation delivers higher yields and cleaner preps. Wiegmans1 antoine leimgruber3 4 and andreas mo ller1 2 1tumour microenvironment laboratory qimr berghofer medical research institute herston qld australia.

For 30 minutes to remove cells and debris. Other kits and. Leveraging our years of exosome expertise the sbi team has pushed exoquick tc extracellular vesicle ev isolation technology to new peaks of performance with exoquick tc ultra for tissue culture media and most biofluids. Isolation of exosomes can be a tedious non specific and difficult process.

Total exosome isolation reagent from cell culture media enables simple and reliable concentration of intact exosomes from cell media samples with a protocol that is scalable depending on your sample size.