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Abstract: PO0297

Human Mesenchymal Stem Cells Cultured in a Hollow Fiber Bioreactor Maintain Constant Levels of Exosomes in the Perfusion Medium: Relevance to the Simultaneous Production of Two Biotherapeutic Agents

Session Information

  • Bioengineering
    October 22, 2020 | Location: On-Demand
    Abstract Time: 10:00 AM - 12:00 PM

Category: Bioengineering

  • 300 Bioengineering

Authors

  • Westenfelder, Christof, SymbioCellTech, LLC, Salt Lake City, Utah, United States
  • Gooch, Anna, SymbioCellTech, LLC, Salt Lake City, Utah, United States
  • Skliar, Mikhail, University of Utah, Salt Lake City, Utah, United States
  • Chowdhury, Sabiha Sultana, SymbioCellTech, LLC, Salt Lake City, Utah, United States
  • Zhang, Ping, SymbioCellTech, LLC, Salt Lake City, Utah, United States
  • Hu, Zhuma, SymbioCellTech, LLC, Salt Lake City, Utah, United States
Background

We have shown that the administration of allogeneic Mesenchymal Stem Cells (MSC) to patients at high risk for Acute Kidney Injury (AKI) following on-pump cardiac surgery prevents AKI and progression to Chronic Kidney Disease (CKD).
Treatment of rats with severe, progressive IRI AKI with MSC-derived exosomes affords significant survival benefits and rescues their renal function (see abstract this meeting). The current study examined the possibility to simultaneously collect MSC-derived exosomes while culturing human MSCs, both used for various therapies in renal and other diseases. This approach, if successful, would
be cost saving, efficient and facilitate up-scaling of the production of both MSCs and their exosomes.

Methods

Human MSCs (20x10e6) were loaded into a hollow fiber Cell Expansion System (Quantum®, TERUMOBct; pre-conditioned for cell adhesion with Fibronectin) and expanded using αMEM with 5% human Platelet Lysate (hPL). The number of exosomes in aliquots of the perfusion medium were monitored (NanoSight instrument) throughout the course of cell expansion.

Results

MSCs reached ~ 90% confluence within 12 days, yielding 500x10e6 MSCs.
The number of exosomes/nanoparticles derived from the 5% hPL per se was 4±1x10e11/mL. Post seeding of MSCs in the bioreactor, exosome numbers in the perfusate decreased and stabilized at 1-1.5x10e11/mL. The size of collected exosomes was between 60 and 100 nm.

Conclusion

The data from this pilot study demonstrate that hPL-derived exosomes or nanoparticles are taken up by the expanding MSCs, which lowers their total number in the perfusion medium. However, exsome numbers stabilized during the subsequent cell expansion, indicating that growing MSCs release high numbers of exosomes. This conclusion will be confirmed by speciating hPL- and MSC-derived exosomes, using specific markers for each type of nanoparticle. Together, these observations show promise for the efficient generation of MSCs and their exosomes to be used for various clinical applications.

Funding

  • Commercial Support