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Editorial
. 2021 Jan;10(3):e12061.
doi: 10.1002/jev2.12061. Epub 2021 Jan 25.

Foetal bovine serum influence on in vitro extracellular vesicle analyses

Brandon M Lehrich  1 , Yaxuan Liang  2 , Massimo S Fiandaca  3
Affiliations
Editorial

Foetal bovine serum influence on in vitro extracellular vesicle analyses

Brandon M Lehrich et al. J Extracell Vesicles. 2021 Jan.
No abstract available

Keywords: EV‐depleted fbs; cell culture media; exosomes; extracellular vesicles; foetal bovine serum; in vitro; serum‐free media.

PubMed Disclaimer

Conflict of interest statement

The authors report no conflict of interest concerning the materials or methods used in this study or the findings specified in this paper.

Figures

FIGURE 1
FIGURE 1
Illustration of FBS‐derived EV and other aggregated non‐EV nanoparticle contamination in conditioned culture media. Specifically, FBS supplies exogenously provided EVs and non‐vesicular contaminants. Adapted from Lehrich et al. Int J Mol Sci, 2018. 19(11)
FIGURE 2
FIGURE 2
FBS EV‐depletion Protocol Efficiencies. (A) FBS‐derived EV particle depletion performed at different time points quantified using NTA with a NanoSight NS‐500 instrument (n = 3 replicates). UC spin occurred at 120,000 x g following a 10,000 x g initial spin for 40 min. Reproduced from Eitan et al. J Extracell Vesicles, 2015. 4: p. 26373. (B) FBS exRNA remaining following depletion over time. Reproduced from Shelke et al. J Extracell Vesicles, 2014. 3. (C) FBS exRNA remaining in supernatant following UC depletion over time. Reproduced from Wei et al. Sci Rep, 2016. 6: p. 31175. (D) Presence of small EVs was determined by transmission electron microscopy. Vesicles are from the pellet preparation of EV‐depleted FBS following an 18‐h UC. Scale bar is 200 nm. Reproduced from Shelke et al. J Extracell Vesicles, 2014. 3. (E) Native FBS, UC EV‐depleted FBS media, and Exo‐FBS™ particle size distributions. Reproduced from Lehrich et al. Int J Mol Sci, 2018. 19(11). (F) Native FBS, UC EV‐depleted FBS media, and Exo‐FBS™ particle concentration differences. Reproduced from Lehrich et al. Int J Mol Sci, 2018. 19(11)
FIGURE 3
FIGURE 3
Extracellular RNA Existence within EV‐depleted FBS Media. (A) Heatmap demonstrating miRNAs and small noncoding RNAs that were abundant in different FBS media conditions (native, Exo‐FBS™, ultracentrifugation, and ultrafiltration). Reproduced from Mannerström et al. Sci Rep, 2019. 9(1): p. 5538. (B) Top; RNA profiles in different FBS conditions from the EVs isolated from the retentate. Bottom; RNA profiles in the FBS media preparations utilized. Reproduced from Kornilov et al. J Extracell Vesicles, 2018. 7(1): p. 1422674. FBS transcript sequence counts mapped to (C) bovine and (D) human reference genomes for varying RNA species in each sample (native, Exo‐FBS™, ultracentrifugation, and ultrafiltration). Reproduced from Mannerström et al. Sci Rep, 2019. 9(1): p. 5538. (E) The Euler diagram provides visual representation of the total quantity of distinct RNAs per media condition. Reproduced from Mannerström et al. Sci Rep, 2019. 9(1): p. 5538
FIGURE 4
FIGURE 4
Deterred Cell Growth in EV‐depleted FBS Media. (A) Human primary myoblasts cultured in native or EV‐depleted FBS media. Reproduced from Aswad et al. BMC Biotechnol, 2016. 16: p. 32. (B) Mouse primary astrocytes cultured in native or EV‐depleted FBS media (via ultracentrifugation or Exo‐FBS™). Reproduced from Lehrich et al. Int J Mol Sci, 2018. 19(11). (C) Illustration that addition of FBS‐derived EVs to EV‐depleted FBS media stimulates migratory cell phenotype in an epithelial cell line. Reproduced from Shelke et al. J Extracell Vesicles, 2014. (D) Cellular phenotypic changes in HIV‐1 infected H9 and PM1 cell cultures under different FBS media conditions (replete = native; TF‐EVD = EV‐depleted FBS media). Adapted from Liao et al. Sci Rep, 2017. 7(1): p. 2558
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