Abstract
Mesenchymal stromal cell (MSC) therapies have been pursued for a broad spectrum of
indications but mixed reports on clinical efficacy have given rise to some degree
of skepticism regarding the effectiveness of this approach. However, recent reports
of successful clinical outcomes and regulatory approvals for graft-versus-host disease,
Crohn's disease and critical limb ischemia have prompted a shift in this perspective.
With hundreds of clinical trials involving MSCs currently underway and an increasing
demand for large-scale manufacturing protocols, there is a critical need to develop
standards that can be applied to processing methods and to establish consensus assays
for both MSC processing control and MSC product release. Reference materials and validated,
uniformly applied tests for quality control of MSC products are needed. Here, we review
recent developments in MSC manufacturing technologies, release testing and potency
assays. We conclude that, although MSCs hold considerable promise clinically, economies
of scale have yet to be achieved although numerous bioreactor technologies for scalable
production of MSCs exist. Additionally, rigorous disease-specific product testing
and comprehensive understanding of mechanisms of action, which are linked to relevant
process and product release potency assays, will be required to ensure that these
therapies continue to be successful.
Key Words
To read this article in full you will need to make a payment
Purchase one-time access:
Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online accessOne-time access price info
- For academic or personal research use, select 'Academic and Personal'
- For corporate R&D use, select 'Corporate R&D Professionals'
Subscribe:
Subscribe to CytotherapyAlready a print subscriber? Claim online access
Already an online subscriber? Sign in
Register: Create an account
Institutional Access: Sign in to ScienceDirect
References
- Stem cell therapies in clinical trials: progress and challenges.Cell Stem Cell. 2015; 17: 11-22
- Discrepancies in autologous bone marrow stem cell trials and enhancement of ejection fraction (DAMASCENE): weighted regression and meta-analysis.BMJ: British Medical Journal. 2014; : 348
- Expanded allogeneic adipose-derived mesenchymal stem cells (Cx601) for complex perianal fistulas in Crohn's disease: a phase 3 randomised, double-blind controlled trial.Lancet. 2016; 388: 1281-1290
- First approval of regenerative medical products under the PMD act in Japan.Cell Stem Cell. 2016; 18: 434-435
Stempeutics [accessed 08.18]. Available from: http://www.stempeutics.com/stempeucel.html.
- The development of fibroblast colonies in monolayer cultures of guinea-pig bone marrow and spleen cells.Cell Tissue Kinet. 1970; 3: 393-403
- Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement.Cytotherapy. 2006; 8: 315-317
- Stromal cells from the adipose tissue-derived stromal vascular fraction and culture expanded adipose tissue-derived stromal/stem cells: a joint statement of the International Federation for Adipose Therapeutics and Science (IFATS) and the International Society for Cellular Therapy (ISCT).Cytotherapy. 2013; 15: 641-648
- Peak MSC—are we there yet?.Frontiers in Medicine. 2018; 5 (14 pages)
- Mesenchymal stem cells for regenerative therapy: optimization of cell preparation protocols.Biomed Res Int. 2014; 2014951512
- UC blood-derived mesenchymal stromal cells: an overview.Cytotherapy. 2007; 9: 717-726
- Manufacturing human mesenchymal stem cells at clinical scale: process and regulatory challenges.Appl Microbiol Biotechnol. 2018; 102: 3981-3994
- Chapter 5 - Mesenchymal stromal cell production in academic centers: challenges and opportunities.in: Viswanathan S Hematti P Mesenchymal Stromal Cells. Academic Press, Boston2017: 121-138
- Chapter 6 - Bioreactor for scale-up: process control.Mesenchymal Stromal Cells. 2017; : 139-178
- Concise review: process development considerations for cell therapy.Stem Cells Transl Med. 2015; 4: 1155-1163
- Commercial scale manufacturing of allogeneic cell therapy.Front Med (Lausanne). 2018; 5: 233
- Concise review: guidance in developing commercializable autologous/patient-specific cell therapy manufacturing.Stem Cells Transl Med. 2013; 2: 871-883
Millicell® HY Multilayer Culture Flasks: Millipore Sigma; [accessed 09.18]. Available from: http://www.emdmillipore.com/CA/en/product/Millicell-HY-Multilayer-Culture-Flasks,MM_NF-C85149.
BD Falcon™ Cell Culture Multi-Flask: BD Biosciences; [accessed 09.18]. Available from: http://www.bdbiosciences.com/documents/multiflasks-data-sheet.pdf.
Corning™ Falcon™ Cell Culture Multi Flasks Fisher Scientific; [accessed 09.18]. Available from: https://www.fishersci.ca/shop/products/falcon-cell-culture-multi-flasks-2/p-4254206.
- Meeting lot-size challenges of manufacturing adherent cells for therapy: BioProcess International.([accessed 08.18]. Available from:)
- Annual Report: Osiris Therapeutics.([accessed 08.18]. Available from:)
NANT 001 System: VivaBioCell; [accessed 09.18]. Available from: http://www.vivabiocell.it/nant001/.
Xpansion® Multiplate Bioreactor System: PALL; [accessed 09.18]. Available from: https://shop.pall.com/us/en/biotech/cell-culture/bioreactors/xpansion-multiplate-bioreactor-system-zidhw7uq21i.
- Production process for stem cell based therapeutic implants: expansion of the production cell line and cultivation of encapsulated cells.Adv Biochem Eng Biotechnol. 2010; 123: 143-162
- Packed bed bioreactor for the isolation and expansion of placental-derived mesenchymal stromal cells.PLoS One. 2015; 10e0144941
- Technologies for large-scale umbilical cord-derived MSC expansion: Experimental performance and cost of goods analysis.Biochem Eng J. 2018; 135: 36-48
- Interest in hollow-fiber perfusion bioreactors is growing: BioProcess International.([accessed 08.18]. Available from:)
- Pros and cons of different expansion systems for manufacturing of an allogeneic cell therapy: Regenesys.([accessed 09.18]. Available from:)
- Efficient manufacturing of therapeutic mesenchymal stromal cells with the use of the Quantum Cell Expansion System.Cytotherapy. 2014; 16: 1048-1058
Single-Use Bioreactors: ThermoFisher Scientific; [accessed 09.18]. Available from: https://www.thermofisher.com/ca/en/home/life-science/bioproduction/single-use-bioprocessing/single-use-equipment/single-use-bioreactors.html.
BioBLU® c Single-Use Vessels: Eppendorf; [accessed 09.18]. Available from: https://online-shop.eppendorf.ca/CA-en/Bioprocess-44559/Vessels-66270/BioBLUc-Single-Use-Vessels-PF-157954.html.
Allegro™ Single-Use Stirred Tank Bioreactors: PALL Corporation; [accessed 09.18]. Available from: https://biotech.pall.com/en/landing/stirred-tank-bioreactors.html?utm_source=google&utm_medium=cpc&utm_content=website-conversions&utm_term=%2Bstirred%20%2Bbioreactor&gclid=CjwKCAjwlejcBRAdEiwAAbj6KeW5hjnftF3ghpzswdMx976lOs6OjMXWwFR0Z0zY6t-xUqCqDwhpjhoCUjwQAvD_BwE.
- Process development for expansion of human mesenchymal stromal cells in a 50L single-use stirred tank bioreactor.Biochem Eng J. 2017; 120: 49-62
- Systematic microcarrier screening and agitated culture conditions improves human mesenchymal stem cell yield in bioreactors.Biotechnol J. 2016; 11: 473-486
- Microcarrier choice and bead-to-bead transfer for human mesenchymal stem cells in serum-containing and chemically defined media.Process Biochem. 2017; 59: 255-265
- Bioreactor manufacturing platforms for cell therapies: BioProcess International.([accessed 08.18]. Available from:)
- Biodegradable poly-epsilon-caprolactone microcarriers for efficient production of human mesenchymal stromal cells and secreted cytokines in batch and fed-batch bioreactors.Cytotherapy. 2017; 19: 419-432
- The local structure of turbulence in incompressible viscous fluid for very large Reynolds numbers.Proceedings of the Royal Society of London Series A: Mathematical and Physical Sciences. 1991; 434: 9-13
- Hydrodynamic effects on animal cells grown in microcarrier cultures.Biotechnol Bioeng. 2006; 95: 295-305
Innovative Vertical-Wheel™ Technology: PBS Biotech; [accessed 09.18]. Available from: https://www.pbsbiotech.com/vertical-wheel.html.
- Production of oncolytic adenovirus and human mesenchymal stem cells in a single-use, Vertical-Wheel bioreactor system: Impact of bioreactor design on performance of microcarrier-based cell culture processes.Biotechnol Prog. 2015; 31: 1600-1612
- Biodistribution of mesenchymal stem/stromal cells in a preclinical setting.Stem Cells Int. 2013; 2013678063
- Validation of the microbiological testing of tissue preparations using the BACTEC blood culture system.Transfus Med Hemother. 2012; 39: 387-390
- Roadmap to approval: use of an automated sterility test method as a lot release test for Carticel® autologous cultured chondrocytes.Cytotherapy. 2005; 7: 531-541
- Guidance, Compliance & Regulatory Information (Biologics).U.S. Food and Drug Administration, 2018 ([accessed 08.18]. Available from:)
- Long-term cultures of bone marrow-derived human mesenchymal stem cells frequently undergo spontaneous malignant transformation.Cancer Res. 2009; 69: 5331-5339
- Spontaneous human adult stem cell transformation.Cancer Res. 2005; 65: 3035-3039
- Retraction: Spontaneous human adult stem cell transformation.Cancer Res. 2010; 70: 6682
- Spontaneous malignant transformation of human mesenchymal stem cells reflects cross-contamination: putting the research field on track – Letter.Cancer Res. 2010; 70: 6393-6396
- Defining the risks of mesenchymal stromal cell therapy.Cytotherapy. 2010; 12: 576-578
- Risk of tumorigenicity in mesenchymal stromal cell-based therapies–bridging scientific observations and regulatory viewpoints.Cytotherapy. 2013; 15: 753-759
- Clinical-grade production of human mesenchymal stromal cells: occurrence of aneuploidy without transformation.Blood. 2010; 115: 1549-1553
- Long-term cultured mesenchymal stem cells frequently develop genomic mutations but do not undergo malignant transformation.Cell Death Dis. 2013; 4: e950
- Growth kinetics, self-renewal, and the osteogenic potential of purified human mesenchymal stem cells during extensive subcultivation and following cryopreservation.J Cell Biochem. 1997; 64: 278-294
- Proliferation kinetics and differentiation potential of ex vivo expanded human bone marrow stromal cells.Exp Hematol. 2000; 28: 707-715
- Comparison of trypan blue dye exclusion and fluorometric assays for mammalian cell viability determinations.Biotechnol Prog. 1993; 9: 671-674
Automated cell counter NucleoCounter® NC-200™: Chemometec; [accessed 09.18]. Available from: https://chemometec.com/cell-counters/cell-counter-nc-200-nucleocounter/.
Vi-CELL XR Cell Counter for Cell Viability: Beckman Coulter; [accessed 09.18]. Available from: http://bc-particle.com/?page_id=46/?pi_ad_id=217813475218.
Automated Cell Counters: ThermoFisher Scientific; [accessed 09.18]. Available from: https://www.thermofisher.com/ca/en/home/life-science/cell-analysis/cell-analysis-instruments/automated-cell-counters.html.
- Gene activation of the apoptotic caspase cascade following cryogenic storage.Cell Preserv Technol. 2002; 1: 63-80
- Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis.Blood. 1994; 84: 1415-1420
- A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V.J Immunol Methods. 1995; 184: 39-51
- Identification of stromal cell precursors in human bone marrow by a novel monoclonal antibody, STRO-1.Blood. 1991; 78: 55-62
- Cell surface antigens on human marrow-derived mesenchymal cells are detected by monoclonal antibodies.Bone. 1992; 13: 69-80
- The monoclonal antibody SH-2, raised against human mesenchymal stem cells, recognizes an epitope on endoglin (CD105).Biochem Biophys Res Commun. 1999; 265: 134-139
- The SH-3 and SH-4 Antibodies recognize distinct epitopes on CD73 from human mesenchymal stem cells.Biochem Biophys Res Commun. 2001; 289: 519-524
- Minimal criteria for defining multipotent mesenchymal stromal cells.The International Society for Cellular Therapy position statement. Cytotherapy. 2006; 8: 315-317
- A novel monoclonal antibody (STRO-3) identifies an isoform of tissue nonspecific alkaline phosphatase expressed by multipotent bone marrow stromal stem cells.Stem Cells Dev. 2007; 16: 953-964
- Heat shock protein-90 beta is expressed at the surface of multipotential mesenchymal precursor cells: generation of a novel monoclonal antibody, STRO-4, with specificity for mesenchymal precursor cells from human and ovine tissues.Stem Cells Dev. 2009; 18: 1253-1262
- Isolation of bone marrow mesenchymal stem cells by anti-nerve growth factor receptor antibodies.Exp Hematol. 2002; 30: 783-791
- Self-renewing osteoprogenitors in bone marrow sinusoids can organize a hematopoietic microenvironment.Cell. 2007; 131: 324-336
- SSEA-4 identifies mesenchymal stem cells from bone marrow.Blood. 2007; 109: 1743-1751
- Phenotypic characterization of distinct human bone marrow-derived MSC subsets.Ann N Y Acad Sci. 2009; 1176: 124-134
- Mesenchymal stem cells: immune evasive, not immune privileged.Nat Biotechnol. 2014; 32: 252-260
- Immunological characteristics of human mesenchymal stem cells and multipotent adult progenitor cells.Immunol Cell Biol. 2013; 91: 32-39
- Immunoprivileged no more: measuring the immunogenicity of allogeneic adult mesenchymal stem cells.Stem Cell Res Ther. 2017; 8: 288
- Changes in phenotype and differentiation potential of human mesenchymal stem cells aging in vitro.Stem Cell Res Ther. 2018; 9: 131
- MSC-based product characterization for clinical trials: an FDA perspective.Cell Stem Cell. 2014; 14: 141-145
- 21CFR600.3 BIOLOGICAL PRODUCTS: GENERAL.U.S. Food and Drug Administration, 2018 ([accessed 09.18]. Available from:)
- A study on ensuring the quality and safety of pharmaceuticals and medical devices derived from the processing of autologous human somatic stem cells.Regen Ther. 2015; 2: 57-69
- Study of the stability of packaging and storage conditions of human mesenchymal stem cell for intra-arterial clinical application in patient with critical limb ischemia.Eur J Pharm Biopharm. 2014; 86: 459-468
- Effect of holding time, temperature and different parenteral solutions on viability and functionality of adult bone marrow-derived mesenchymal stem cells before transplantation.J Tissue Eng Regen Med. 2008; 2: 436-444
- Duration of in vitro storage affects the key stem cell features of human bone marrow-derived mesenchymal stromal cells for clinical transplantation.Cytotherapy. 2013; 15: 460-466
- Cryopreserved mesenchymal stromal cells display impaired immunosuppressive properties as a result of heat-shock response and impaired interferon-gamma licensing.Cytotherapy. 2012; 14: 147-152
- Mechanisms of mesenchymal stem/stromal cell function.Stem Cell Res Ther. 2016; 7: 125
- International Society for Cellular Therapy perspective on immune functional assays for mesenchymal stromal cells as potency release criterion for advanced phase clinical trials.Cytotherapy. 2016; 18: 151-159
- Soliciting strategies for developing cell-based reference materials to advance mesenchymal stromal cell research and clinical translation.Stem Cells Dev. 2014; 23: 1157-1167
- Comparing the immunomodulatory properties of bone marrow, adipose tissue, and birth-associated tissue mesenchymal stromal cells.Front Immunol. 2015; 6: 560
- Mesenchymal stem cells and immunomodulation: current status and future prospects.Cell Death Dis. 2016; 7: e2062
- Immunoregulation by mesenchymal stem cells: biological aspects and clinical applications.J Immunol Res. 2015; 2015394917
- Mesenchymal stem cells in tissue repair.Front Immunol. 2013; 4: 201
- Human mesenchymal stem cells (MSCs) for treatment towards immune- and inflammation-mediated diseases: review of current clinical trials.J Biomed Sci. 2016; 23: 76
- Immunoregulatory mechanisms of mesenchymal stem and stromal cells in inflammatory diseases.Nat Rev Nephrol. 2018; 14: 493-507
- Potency analysis of mesenchymal stromal cells using a combinatorial assay matrix approach.Cell Rep. 2018; 22: 2504-2517
- Mesenchymal stem cells inhibit and stimulate mixed lymphocyte cultures and mitogenic responses independently of the major histocompatibility complex.Scand J Immunol. 2003; 57: 11-20
- A reproducible immunopotency assay to measure mesenchymal stromal cell-mediated T-cell suppression.Cytotherapy. 2015; 17: 140-151
- Regulatory perspective on in vitro potency assays for human mesenchymal stromal cells used in immunotherapy.Cytotherapy. 2017; 19: 784-797
- Therapeutic delivery specifications identified through compartmental analysis of a mesenchymal stromal cell-immune reaction.Sci Rep. 2018; 8: 6816
- Long-term complications, immunologic effects, and role of passage for outcome in mesenchymal stromal cell therapy.Biol Blood Marrow Transplant. 2012; 18: 557-564
- Apoptosis in mesenchymal stromal cells induces in vivo recipient-mediated immunomodulation.Sci Transl Med. 2017; 9
- Mesenchymal stem cells promote macrophage polarization toward M2b-like cells.Exp Cell Res. 2016; 348: 36-45
- Therapeutic potential of adipose-derived stem cells and macrophages for ischemic skeletal muscle repair.Regen Med. 2017; 12: 153-167
- Human macrophage regulation via interaction with cardiac adipose tissue-derived mesenchymal stromal cells.J Cardiovasc Pharmacol Ther. 2013; 18: 78-86
- Development of a flow cytometry-based potency assay for measuring the in vitro immunomodulatory properties of mesenchymal stromal cells.Immunol Lett. 2016; 177: 38-46
- In Vitro Macrophage Phagocytosis Assay.Methods Mol Biol. 2015; 1339: 235-246
- Allogeneic human mesenchymal stem cell therapy (remestemcel-L, Prochymal®) as a rescue agent for severe treatment resistant graft-versus-host disease in pediatric patients.Biol Blood Marrow Transplant. 2014; 20: 229-235
- Prochymal improves response rates in patients with steroid-refractory acute graft versus host disease (SR-GVHD) involving the liver and gut: results of a randomized, placebo-controlled, multicenter phase III trial in GVHD.Biol Blood Marrow Transplant. 2010; 16: S169-SS70
- Adult human mesenchymal stem cells added to corticosteroid therapy for the treatment of acute graft-versus-host disease.Biol Blood Marrow Transplant. 2009; 15: 804-811
- Inducible indoleamine 2,3-dioxygenase 1 and programmed death ligand 1 expression as the potency marker for mesenchymal stromal cells.Cytotherapy. 2018; 20: 639-649
- TSG-6 as a biomarker to predict efficacy of human mesenchymal stem/progenitor cells (hMSCs) in modulating sterile inflammation in vivo.Proc Natl Acad Sci U S A. 2014; 111: 16766-16771
- Aggregation of human mesenchymal stromal cells (MSCs) into 3D spheroids enhances their antiinflammatory properties.Proc Natl Acad Sci U S A. 2010; 107: 13724-13729
- Prostaglandin E2 indicates therapeutic efficacy of mesenchymal stem cells in experimental traumatic brain injury.Stem Cells. 2017; 35: 1416-1430
- Immunosuppressive capacity of mesenchymal stem cells correlates with metabolic activity and can be enhanced by valproic acid.Stem Cell Res Ther. 2017; 8: 100
- Morphological features of IFN-gamma-stimulated mesenchymal stromal cells predict overall immunosuppressive capacity.Proc Natl Acad Sci U S A. 2017; 114: E2598-Ee607
Cell X Technologies [accessed 09.18]. Available from: https://www.cellxtechnologies.com/.
- The angiogenic properties of mesenchymal stem/stromal cells and their therapeutic potential.Br Med Bull. 2013; 108: 25-53
- Proangiogenic features of mesenchymal stem cells and their therapeutic applications.Stem Cells Int. 2016; 20161314709
- Bioenergetic and functional consequences of bone marrow-derived multipotent progenitor cell transplantation in hearts with postinfarction left ventricular remodeling.Circulation. 2007; 115: 1866-1875
- Molecular imaging of the paracrine proangiogenic effects of progenitor cell therapy in limb ischemia.Circulation. 2013; 127: 710-719
- Development of a surrogate angiogenic potency assay for clinical-grade stem cell production.Cytotherapy. 2012; 14: 994-1004
- Development of a surrogate potency assay to determine the angiogenic activity of Stempeucel(R), a pooled, ex-vivo expanded, allogeneic human bone marrow mesenchymal stromal cell product.Stem Cell Res Ther. 2017; 8: 47
- The aortic ring co-culture assay: a convenient tool to assess the angiogenic potential of mesenchymal stromal cells in vitro.J Vis Exp. 2017; (12 pages)
- Assessing angiogenic responses induced by primary human prostate stromal cells in a three-dimensional fibrin matrix assay.Oncotarget. 2016; 7: 71298-71308
- A clinical indications prediction scale based on TWIST1 for human mesenchymal stem cells.EBioMedicine. 2016; 4: 62-73
- Mesenchymal stem cells derived exosomes and microparticles protect cartilage and bone from degradation in osteoarthritis.Sci Rep. 2017; 7: 16214
- Exosome secreted by MSC reduces myocardial ischemia/reperfusion injury.Stem Cell Res. 2010; 4: 214-222
- MSC-derived extracellular vesicles attenuate immune responses in two autoimmune murine models: Type 1 diabetes and uveoretinitis.Stem Cell Reports. 2017; 8: 1214-1225
- Mitochondrial transfer from bone-marrow-derived stromal cells to pulmonary alveoli protects against acute lung injury.Nat Med. 2012; 18: 759-765
- Mitochondrial transfer of mesenchymal stem cells effectively protects corneal epithelial cells from mitochondrial damage.Cell Death Dis. 2016; 7: e2467
- Mesenchymal stromal cells modulate macrophages in clinically relevant lung injury models by extracellular vesicle mitochondrial transfer.Am J Respir Crit Care Med. 2017; 196: 1275-1286
- Intercellular mitochondria trafficking highlighting the dual role of mesenchymal stem cells as both sensors and rescuers of tissue injury.Cell Cycle (Georgetown, Tex). 2018; 17: 712-721
- Concise review: MicroRNA function in multipotent mesenchymal stromal cells.Stem Cells. 2014; 32: 1074-1082
- microRNA-378 promotes mesenchymal stem cell survival and vascularization under hypoxic–ischemic conditions in vitro.Stem Cell Res Ther. 2014; 5: 130
- Mesenchymal stem cells deliver exogenous MicroRNA-let7c via exosomes to attenuate renal fibrosis.Mol Ther. 2016; 24: 1290-1301
Article info
Publication history
Published online: December 06, 2018
Accepted:
October 26,
2018
Received:
September 15,
2018
Identification
Copyright
© 2018 International Society for Cell and Gene Therapy. Published by Elsevier Inc. All rights reserved.
