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EV-Up™ Exosomes Production Medium for MSC (animal component-free)

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EV-Up™ media is Exosomes (EVs) production media for mesenchymal stem cells (MSC). This media is adjusted as complete medium by adding "EV-Up™ MSC EV Production Supplement, AF" to "EV-Up™ EV-Production Basal Medium for MSC, AF". This is serum- and animal-free and applicable to any growth media.

Features

  • Better EVs yield than conventional serum containing media
  • Able to produce EVs with high activity
  • Able to maintain MSC viability

Applicable Cells

  • Bone-marrow-derived MSC
  • Adipose-derived MSC
  • Umbilical cord-derived MSC

MSC EVs Production Protocol

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*1: The complete media which “EV-Up™ MSC EV Production Supplement, AF” added to "EV-Up™ EV-Production Medium for MSC, AF".

When culturing on a 6-well plate

  1. 5×103 cells of various MSC seed and culture using Growth media (with or without serum) until 80-90% confluent.
  2. Growth media discard and change to 4mL of EV-Up™ media. Incubates for 3-5 days.
  3. The culture supernatant collect to a tube and centrifuge at 2,000×g for 20 minutes. Collect the culture supernatant of it in a new tube.
    Able to suppress absorption of EVs to a microtube and chip by adding EV-Save™ Extracellular Vesicle Blocking Reagent (Code No. 058-09261) that 1/100 amount of the culture supernatant.
  4. EVs isolate from the supernatant using MagCapture™ Exosome Isolation Kit PS Ver.2 (Code No. 290-84103) etc..

Application Data

Nano Tracking Analysis (NTA)

EVs isolated from various media supernatant by PS affinity method*2 were analyzed with NTA.

*2: EVs are captured by phosphatidylserine (PS)-binding proteins and metal ions, then captured EVs are eluted with a chelating reagent.

  • DMEM + 10% EV depleted FBS

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  • EV-Up™ media

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  • (A) The number of EVs particles

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    *3: Exosome-Depleted FBS

  • (B) EVs diameter

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MSC cultured in EV-Up™ media released 2.6 times more EVs than MSC cultured
in DMEM + 10% EV-depleted FBS. And the EV’s diameters were almost same.

 

Anti-fibrotic Effect

5×107 particles/mL of EVs isolated from various media supernatant by PS affinity method were added to normal human fetal lung-diploid fibroblasts cells (TIG3) that were stimulated TGFβ. And fibrotic marker (Collagen Ⅲ, αSMA) gene expression was quantified by RT-PCR.

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  • Collagen Ⅲ

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  • α-SMA

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Significantly, MSC EVs produced in EV-Up™ media decreased
the gene expression of fibrotic markers such as Collagen Ⅲ (A) and αSMA (B).

 

Cell Viability

After the expansion of human bone marrow-derived MSC in serum containing media, the media was switched with EV-Up™ media and cultured for five days to produce EVs. After that, cell viability was measured.

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MSC that was cultured in EV-Up™ showed high survival rate like when cultured in DMEM + 10% EV.

 

Exosome Marker Protein Expression

EV marker protein expression (CD9, CD63, CD81) of Evs isolated from various media supernatant were confirmed by ELISA.

  • CD9

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  • CD63

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  • CD81

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EV-Up™ media produced EVs that expressed various EV marker protein.

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EV-Up™ Media

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Exosome Blocking Reagent

Isolation/Purification of Exosome

Product content may differ from the actual image due to minor specification changes etc.

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