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Updated: Nov 4, 2025
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Published on: July 26, 2022
Comparative Analysis of MSC-Derived Exosomes Depending on Cell Culture Media for Regenerative Bioactivity
Jun Yong Kim1,2,3, Won-Kyu Rhim1, Hyo Jeong Seo1
1Department of Biomedical Science, CHA University, 335 Pangyo-ro, Bundang-gu, Seongnam-si, Gyeonggi-do, 13488, Republic of Korea.
Background:
In order to produce and isolate the exosome derived from the cell of interests, a serum free environment (starvation) has been essential for excluding the unknown effect from serum-derived exosomes. Recently, serum-free culture media have been developed as a substitute for serum supplemented media so that MSC proliferates with maintaining the original characteristics of the cells in a serum free condition. Due to the different properties of the exosomes representing the states and characteristics of the origin cells, a study is needed to compare the properties of the cell-derived exosomes according to the cell culture media.
Methods:
To compare the cell culture condition on exosomes, human umbilical cord mesenchymal stem cells (UCMSCs) were cultured with two different media, serum containing media, 10% FBS supplemented DMEM (NM) and serum-free chemically defined media, CellCor™ CD MSC (CDM). To remove FBS-derived exosomes from UCMSC cultured with NM, the medium was replaced with FBS-free DMEM for starvation during exosome isolation. The production yield and expression levels of angiogenic and pro-inflammatory factors were compared. And, the subpopulations of exosome were classified depending on the surface properties and loaded cytokines. Finally, the wound healing and angiogenic effects have been evaluated using in vitro assays.
Results:
The UCMSC-derived exosomes under two different cell culture media could be classified into subpopulations according to the surface composition and loaded cytokines. Especially, exosome derived from UCMSC cultured with CDM showed higher expression levels of cytokines related to regenerative bioactivities which resulted in enhanced wound healing and angiogenesis.
Conclusion:
CDM has the advantages to maintain cell proliferation even during the period of exosome isolations and eliminate unknown side effects caused by serum-derived exosomes. Additionally, exosomes derived from UCMSC cultured with CDM show better wound healing and angiogenic effects due to a lot of regeneration-related cytokines and less pro-inflammatory cytokines compared to with NM.
Insights
Serum-free chemically defined media (CDM) enhances exosome production from mesenchymal stem cells, yielding exosomes with superior regenerative and angiogenic properties for improved wound healing.
Area of Science:
- Biotechnology
- Cell Biology
- Exosome Research
Background:
- Exosome isolation traditionally requires serum-free conditions to avoid contamination from serum-derived exosomes.
- Serum-free culture media are emerging as a way to culture cells while preserving their characteristics.
- Comparing exosome properties based on cell culture media is crucial due to exosomes reflecting their origin cells' state.
Purpose of the Study:
- To compare the impact of serum-containing media versus serum-free chemically defined media on exosome production and properties.
- To evaluate the regenerative and angiogenic potential of exosomes derived from cells cultured under different conditions.
Main Methods:
- Human umbilical cord mesenchymal stem cells (UCMSCs) were cultured in either serum-containing DMEM (NM) or serum-free CDM.
- Exosomes were isolated, and their production yield and expression of angiogenic/pro-inflammatory factors were analyzed.
- In vitro assays assessed the wound healing and angiogenic effects of the derived exosomes.
Main Results:
- UCMSC-derived exosomes could be classified into subpopulations based on surface properties and loaded cytokines.
- Exosomes from CDM-cultured UCMSCs exhibited higher levels of regenerative cytokines.
- Exosomes derived from CDM demonstrated enhanced wound healing and angiogenic effects.
Conclusions:
- Chemically defined media (CDM) supports cell proliferation and exosome isolation without serum-derived exosome contamination.
- Exosomes cultured in CDM show superior wound healing and angiogenic potential due to a favorable cytokine profile.
- CDM offers advantages for producing exosomes with enhanced therapeutic bioactivities.
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