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Purpose <p>Extracellular vesicles (EVs) are nanoparticles that mediate cell signaling through miRNAs and bioactive molecules. The therapeutic effects of mesenchymal stromal cells (MSCs) partly arise from their EV secretion. While MSCs are usually cultured with fetal bovine serum (FBS), platelet-rich plasma (PRP) may serve as an alternative, though its impact on EV release remains unclear. This study evaluates EV secretion by adipose-derived MSCs (AD-MSCs) cultured with PRP instead of FBS.</p> Methods <p>AD-MSCs were cultured for 72&#xa0;h in media supplemented with 10% FBS and 2.5% PRP. After 24&#xa0;h without supplementation, EVs were isolated from the supernatant (<i>n</i> = 3 each) by ultracentrifugation (Type 70 Ti Fixed-Angle Rotor).</p> Results <p>Nanoparticle Tracking Analysis (NTA) indicated that it was feasible to isolate similar amounts of EVs from the MSCs culture supernatant when supplemented with 10% FBS compared to supplementation with 2.5% PRP (<i>p</i> &gt; 0.05). In addition, isolated EVs were similar in size in both scenarios, with means between 100 and 200&#xa0;nm (<i>p</i> &gt; 0.05). Transmission Electron Microscopy (TEM) revealed no significant morphological changes between the EVs of the groups supplemented with PRP and FBS. Western blot was positively marked for TSG101 and CD81 proteins.</p> Conclusion <p>Our findings suggest that PRP is a promising xeno-free alternative under the tested conditions, preserving basic EV characteristics and secretion profile, although further validation is required.</p>

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Platelet-rich plasma as a xeno-free alternative to fetal bovine serum for in vitro expansion of adipose-derived mesenchymal stromal cells and isolation of extracellular vesicles

  • 期刊论文

  • 发表日期：

    2026年6月3日

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  • Ana Carolina Borges Campos,
  • Anne Moraes de Monção,
  • Natália Ferreira Haddad,
  • Almir Jordão da Silva-Junior,
  • Camila Hubner Costabile Wendt,
  • Ana Célia Sá de Carvalho,
  • Tathiana Proença Pampolha,
  • Leandra Santos Baptista,
  • Ana Luiza Frango de Sant’Anna,
  • Denise de Abreu Pereira,
  • José Marques de Brito Neto,
  • Rosalia Mendez-Otero,
  • Juliana Ferreira Vasques,
  • Ronaldo José Farias Corrêa do Amaral

摘要

Purpose

Extracellular vesicles (EVs) are nanoparticles that mediate cell signaling through miRNAs and bioactive molecules. The therapeutic effects of mesenchymal stromal cells (MSCs) partly arise from their EV secretion. While MSCs are usually cultured with fetal bovine serum (FBS), platelet-rich plasma (PRP) may serve as an alternative, though its impact on EV release remains unclear. This study evaluates EV secretion by adipose-derived MSCs (AD-MSCs) cultured with PRP instead of FBS.

Methods

AD-MSCs were cultured for 72 h in media supplemented with 10% FBS and 2.5% PRP. After 24 h without supplementation, EVs were isolated from the supernatant (n = 3 each) by ultracentrifugation (Type 70 Ti Fixed-Angle Rotor).

Results

Nanoparticle Tracking Analysis (NTA) indicated that it was feasible to isolate similar amounts of EVs from the MSCs culture supernatant when supplemented with 10% FBS compared to supplementation with 2.5% PRP (p > 0.05). In addition, isolated EVs were similar in size in both scenarios, with means between 100 and 200 nm (p > 0.05). Transmission Electron Microscopy (TEM) revealed no significant morphological changes between the EVs of the groups supplemented with PRP and FBS. Western blot was positively marked for TSG101 and CD81 proteins.

Conclusion

Our findings suggest that PRP is a promising xeno-free alternative under the tested conditions, preserving basic EV characteristics and secretion profile, although further validation is required.