Mechanical fibrinogen‑depletion supports heparin‑free mesenchymal stem cell propagation in human platelet lysate

Mesenchymal stem cells (MSCs) are often expanded outside the body for clinical studies using pooled human platelet lysate (pHPL). This study addresses the issue of clot formation in pHPL-supplemented medium due to coagulation factors. The study introduces a method to deplete fibrinogen from pHPL by clotting the pHPL in the medium, mechanically disrupting the hydrogel, and removing the resulting fibrin pellet. This process allows for heparin-free MSC propagation. The method facilitates a standardized and GMP-grade generation of medium for future clinical studies by providing entirely humanized cell culture media with high medium recovery rates and low remaining fibrinogen amounts.

• 1
  The clotting procedure reduced fibrinogen levels more than 1000-fold, with a high volume recovery of 99.5%.
• 2
  Enhanced proliferation of MSCs was observed in medium lacking fibrinogen but containing heparin, compared to standard medium.
• 3
  This proliferative response to heparin was not detected in MSCs that had prior contact with fibrinogen during the isolation process.