EV-Up™ EV Production Basal Medium/Supplement for MSC
EV-Up™ EV Production Basal Medium for MSC, AF is a culture medium optimized for the production of extracellular vesicle (EV) from mesenchymal stem cell (MSC). The medium is free of animal-derived components and serum. When used together with the separately available EV-Up™ MSC EV Production Supplement, AF, it enhances EV production, including exosomes.
Product Overview
Features
- Increased EV production compared with conventional serum-supplemented media
- Enhanced biological activity of EVs
- High survival rate of MSCs is maintained
Applicable cells
Applicable to a wide range of MSCs, regardless of their species of origin.
Examples of cells
- Bone marrow-derived MSC
- Adipose-derived MSC
- Umbilical cord-derived MSC
Protocol
For extracellular vesicle isolation, we recommend using PS affinity-based methods, such as the MagCapture™ Exosome Isolation Kit PS Ver.2 or the MassivEV™ EV Purification Column PS.
About MSC growth medium
This product is compatible with any growth medium. For EV production from MSCs, we recommend MSCulture™ High Growth Basal Medium/Supplement.
Example of Protocol (6-well plate)
- Seed MSCs at 5 x 103 cells/cm2 and culture in growth medium until they reach 80–90% confluence.
- Remove the growth medium, add 4 mL of EV-Up™ EV Production Basal Medium for MSC, AF with Supplement, and culture for 3–5 days.
- Collect the cell culture supernatant and centrifuge at 2,000 × g for 20 minutes. Carefully transfer the supernatant to a new tube without disturbing the pellet.
[Note] Adding EV-Save™ Extracellular Vesicle Blocking Reagent at 1/100 of the culture supernatant volume can reduce adsorption of EVs to tubes and pipette tips.
- Purify EVs from the culture supernatant using the PS affinity method or other suitable techniques.
Data
Comparison of EV Particle Counts
EVs were purified from the cell culture supernatant of human bone marrow-derived MSCs cultured in each medium using the PS affinity method and quantified by nanoparticle tracking analysis (NTA) with a NanoSight instrument.
[Result]
The number of EV particles released from MSCs cultured in EV-Up™ was approximately 2.6-fold higher than that from MSCs cultured in conventional medium.
Comparison of EV Marker Protein Expression
EVs were isolated from the cell culture supernatants of human bone marrow–derived MSCs cultured in each medium using the PS affinity method. Levels of EV marker proteins (CD9, CD63, CD81) were measured by ELISA.
[Result]
The cell culture supernatant from EV-Up™ contained higher levels of EV marker proteins than that from conventional medium.
Comparison of Cell Viability
Human bone marrow-derived MSCs were first expanded in a serum-containing medium and then cultured for 5 days in each medium. Cell viability was subsequently measured.
[Result]
MSCs cultured in EV-Up™ exhibited high viability, comparable to that of MSCs cultured in conventional medium.
Antifibrotic Activity of EVs Produced in Different Media
EVs (5 x 108 particles/mL) were purified from the cell culture supernatant of human bone marrow-derived MSCs using the PS affinity method. These EVs were then added to human fetal lung fibroblasts (TIG3 cells) stimulated with TGF-β. Antifibrotic activity was evaluated by quantifying mRNA levels of the fibrosis markers collagen III and α-SMA using RT-PCR.
[Result]
EVs produced from MSCs cultured in EV-Up™ exhibited stronger antifibrotic activity compared with those produced in conventional medium.
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EV-Up™ EV Production Basal Medium for MSC, AF
EV-Up™ MSC EV Production Supplement, AF
For research use or further manufacturing use only. Not for use in diagnostic procedures.
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The prices are list prices in Japan.Please contact your local distributor for your retail price in your region.