Purpose
The primary objective of this study was to reproduce and validate the harvest, processing
and storage of peripheral blood stem cells for a subsequent cartilage repair trial,
evaluating safety, reliability, and potential to produce viable, sterile stem cells.
Methods
Ten healthy subjects (aged 19-44 years) received 3 consecutive daily doses of filgrastim
followed by an apheresis harvest of mononuclear cells on a fourth day. In a clean
room, the apheresis product was prepared for cryopreservation and processed into 4
mL aliquots. Sterility and qualification testing were performed pre-processing and
post-processing at multiple time points out to 2 years. Eight samples were shipped
internationally to validate cell transport potential. One sample from all participants
was cultured to test proliferative potential with colony forming unit (CFU) assay.
Five samples, from 5 participants were tested for differentiation potential, including
chondrogenic, adipogenic, osteogenic, endoderm, and ectoderm assays.
Results
Fresh aliquots contained an average of 532.9 ± 166. × 106 total viable cells/4 mL vial and 2.1 ± 1.0 × 106 CD34+ cells/4 mL vial. After processing for cryopreservation, the average cell count
decreased to 331.3 ± 79. × 106 total viable cells /4 mL vial and 1.5 ± 0.7 × 106 CD34+ cells/4 mL vial CD34+ cells. Preprocessing viability averaged 99% and postprocessing
88%. Viability remained constant after cryopreservation at all subsequent time points.
All sterility testing was negative. All samples showed proliferative potential, with
average CFU count 301.4 ± 63.9. All samples were pluripotent.
Conclusions
Peripheral blood stem cells are pluripotent and can be safely harvested/stored with
filgrastim, apheresis, clean-room processing, and cryopreservation. These cells can
be stored for 2 years and shipped without loss of viability.
Clinical Relevance
This method represents an accessible stem cell therapy in development to augment cartilage
repair.
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Article info
Publication history
Published online: April 30, 2021
Accepted:
April 15,
2021
Received:
July 26,
2020
Footnotes
See commentary on page 3357
The authors report the following potential conflicts of interest or sources of funding: Sponsored by KLSMC Stem Cells and supported by grants from City of Gulf Breeze, Florida, and the State of Florida Department of Health. A.W.A. reports grants from Arthrex Inc. and CGG Medical; personal fees from Arthrex, Smith & Nephew, and Bioventus; and other from Arthrex. K-Y.S. reports patent US 8,377,432 B2. Full ICMJE author disclosure forms are available for this article online, as supplementary material.
Identification
Copyright
© 2021 Published by Elsevier on behalf of the Arthroscopy Association of North America
