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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References
- Benefits of small volume and small syringe for bone marrow aspirations of mesenchymal stem cells.Int Orthop. 2013; 37: 2279-2287
- Quantitative assessment of the yield of osteoblastic connective tissue progenitors in bone marrow aspirate from the iliac crest, tibia, and calcaneus.J Bone Joint Surg Am. 2013; 95: 1312-1316
- Aspiration to obtain osteoblast progenitor cells from human bone marrow: The influence of aspiration volume.JBJS. 1997; 79: 1699-1709
- Optimizing dose and scheduling of filgrastim (granulocyte colony-stimulating factor) for mobilization and collection of peripheral blood progenitor cells in normal volunteers.Blood. 1995; 86: 4437-4445
- Safety and efficacy of hematopoietic stem cell collection from mobilized peripheral blood in unrelated volunteers: 12 years of single-center experience in 3928 donors.Blood. 2009; 114: 3757-3763
- Mobilization and collection of peripheral blood progenitor cells for transplantation.Transfus Apher Sci. 2005; 32: 63-72
- Circulating mesenchymal stem cells.Int J Biochem Cell Biol. 2004; 36: 585-597
- Multipotent progenitor cells are present in human peripheral blood.Circ Res. 2009; 104: 1225-1234
- Human granulocyte colony-stimulating factor (G-CSF) stimulates the in vitro and in vivo development but not commitment of primitive multipotential progenitors from transgenic mice expressing the human G-CSF receptor.Blood. 1998; 92: 4632-4640
- Isolation of mesenchymal stem cells from G-CSF-mobilized human peripheral blood using fibrin microbeads.Bone Marrow Transplant. 2006; 37: 967-976
- Peripheral blood mononuclear cells enhance cartilage repair in in vivo osteochondral defect model.PLoS One. 2015; 10e0133937
- Circulating fibrocytes define a new leukocyte subpopulation that mediates tissue repair.Mol Med. 1994; 1: 71-81
- Human circulating CD14+ monocytes as a source of progenitors that exhibit mesenchymal cell differentiation.J Leukoc Biol. 2003; 74: 833-845
- Flt3+ macrophage precursors commit sequentially to osteoclasts, dendritic cells and microglia.BMC Immunol. 2002; 3: 1-11
- R848, a toll-like receptor 7 agonist, inhibits osteoclast differentiation but not survival or bone-resorbing function of mature osteoclasts.Cytotechnology. 2012; 64: 331-339
- Development, differentiation, and maturation of Kupffer cells.Microsc Res Tech. 1998; 39: 350-364
- The macrophage.Bioessays. 1995; 17: 977-986
- Cardiomyogenic potential of mesenchymal progenitors derived from human circulating CD14+ monocytes.Stem Cells Dev. 2006; 14: 676-686
- Repair of large full-thickness cartilage defect by activating endogenous peripharal blood stem cells and autologous periosteum flap transplantation combined with patellofemoral realignment.Knee. 2014; 21: 609-612
- Human peripheral blood derived mesenchymal stem cells demonstrate similar characteristics and chondrogenic differentiation potential to bone marrow derived mesenchymal stem cells.J Orthop Res. 2012; 30: 634-642
- High tibial osteotomy in combination with chondrogenesis after stem cell therapy: A histologic report of 8 cases.Arthroscopy. 2015; 31: 1909-1920
- Articular cartilage regeneration with autologous peripheral blood progenitor cells and hyaluronic acid after arthroscopic subchondral drilling: A report of 5 cases with histology.Arthroscopy. 2011; 27: 493-506
- Articular cartilage regeneration with autologous peripheral blood stem cells versus hyaluronic acid: A randomized controlled trial.Arthroscopy. 2013; 29: 684-694
- Articular cartilage regeneration with autologous marrow aspirate and hyaluronic acid: An experimental study in a goat model.Arthroscopy. 2009; 25: 1391-1400
- Factors affecting mobilization of CD34+ cells in normal donors with filgrastim.Transfusion (Paris). 1997; 37: 507-512
- Peripheral blood stem cell yield in 400 normal donors mobilised with granulocyte colony-stimulating factor (G-CSF): Impact of age, sex, donor weight and type of G-CSF used.Br J Haematol. 2006; 134: 517-525
- Allogeneic blood stem cell transplantation for refractory leukemia and lymphoma: Potential advantage of blood over marrow allografts.Blood. 1995; 85: 1659-1665
- Predictive factors that affect the mobilization of CD34+ cells in healthy donors treated with recombinant granulocyte colony-stimulating factor (G-CSF).J Clin Apher. 2006; 21: 169-175
- Long-term storage of peripheral blood stem cells frozen and stored with a conventional liquid nitrogen technique compared with fresh frozen and stored in a mechanical freezer.Transfusion (Paris). 2010; 50: 808-819
- A new source of mesenchymal stem cells for articular cartilage repair: MSCs derived from mobilized peripheral blood share similar biological characteristics in vitro and chondrogenesis in vivo as MSCs from bone marrow in rabbit model.Am J Sports Med. 2014; 42: 592-601
- Effects of recombinant human granulocyte colony-stimulating factor on hematopoietic progenitor cells in cancer patients.Blood. 1988; 72: 2074-2081
- Follow-up of healthy donors receiving granulocyte colony-stimulating factor for peripheral blood progenitor cell mobilization and collection. Results of the Spanish Donor Registry.Haematologica. 2008; 93: 735-740
- Biologic and clinical effects of granulocyte colony-stimulating factor in normal individuals.Blood. 1996; 88: 2819-2825
- Effects of G-CSF administration and peripheral blood progenitor cell collection in 20 healthy donors.Ann Hematol. 1996; 72: 269-272
- Cartilage lesions of the knee treated with blood mesenchymal stem cells—Results.Ortoped Traumatol Rehabil. 2012; 14: 569-577
- Combination of intra-articular autologous activated peripheral blood stem cells with growth factor addition/preservation and hyaluronic acid in conjunction with arthroscopic microdrilling mesenchymal cell stimulation improves quality of life and regenerates articular cartilage in early osteoarthritic knee disease.J Med Assoc Thai. 2013; 96: 580-588
- The incidence of infectious complications of central venous catheters at the subclavian, internal jugular, and femoral sites in an intensive care unit population.Crit Care Med. 2005; 33: 13-20
- Safety and efficacy of mobility interventions in patients with femoral catheters in the ICU: A prospective observational study.Cardiopulm Phys Ther J. 2013; 24: 12-17
- Peripheral blood progenitor cell mobilization for autologous and allogeneic hematopoietic cell transplantation: guidelines from the American Society for Blood and Marrow Transplantation.Biol Blood Marrow Transplant. 2014; 20: 1262-1273
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
