Manufacturing mesenchymal stromal cells for clinical applications: A survey of Good Manufacturing Practices at U.S. academic centers

  • Donald G. Phinney
    Correspondence: Donald G. Phinney, PhD, Department of Molecular Medicine, The Scripps Research Institute, Scripps Florida, A215, 130 Scripps Way, Jupiter, FL 33458.
    Department of Molecular Medicine, The Scripps Research Institute–Scripps Florida, Jupiter, Florida, USA
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  • Jacques Galipeau
    Department of Medicine and Carbone Cancer Center, University of Wisconsin in Madison, Madison, Wisconsin, USA
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      Background aims

      Mesenchymal stromal cells (MSC) have gained prominence in the field of regenerative medicine due to their excellent safety profile in human patients and recently demonstrated efficacy in late-stage clinical studies. A prerequisite to achieving successful MSC-based therapies is the development of large-scale manufacturing processes that preserve the biological potency of the founder cell population. Because no standardized manufacturing process exists for MSCs, understanding differences in these processes among U.S. academic facilities would allow for better comparison of results obtained in the clinical setting.


      We collected information through a questionnaire sent to U.S. academic centers that produce MSCs under Good Manufacturing Practice conditions.


      The survey provided information on the number and geographic location of academic facilities in the United States and major trends in their manufacturing practices. For example, most facilities employed MSCs enriched from bone marrow by plastic adherence and expanded in media supplemented with pooled human platelet lysate. Sterility testing and product identification via cell surface phenotype analysis were commonly reported practices, whereas initial and working cell plating densities, culture duration, product formulation and the intended use of the MSC product were highly variable among facilities. The survey also revealed that although most facilities assessed product potency, the methods used were limited in scope compared with the broad array of intended clinical applications of the product.


      Survey responses reported herein offer insight into the current best practices used to manufacture MSC-based products in the United States and how these practices may affect product quality and potency. The responses also provide a foundation to establish standardized manufacturing platforms.

      Key Words

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        • Lalu M.M.
        • Mazzarello S.
        • Zlepnig J.
        • Dong Y.Y.R.
        • Montroy J.
        • McIntyre L.
        • et al.
        Safety and efficacy of adult stem cell therapy for acute myocardial infarction and ischemic heart failure (SafeCell Heart): A systematic review and meta-analysis.
        Stem Cells Transl Med. 2018; 7: 857-866
        • Wang L.
        • Zhu C.Y.
        • Ma D.X.
        • Gu Z.Y.
        • Xu C.C.
        • Wang F.Y.
        • et al.
        Efficacy and safety of mesenchymal stromal cells for the prophylaxis of chronic graft-versus-host disease after allogeneic hematopoietic stem cell transplantation: a meta-analysis of randomized controlled trials.
        Ann Hematol. 2018; 97: 1941-1950
        • Galipeau J.
        • Sensebe L.
        Mesenchymal stromal cells: Clinical challenges and therapeutic opportunities.
        Cell Stem Cell. 2018; 22: 824-833
        • Liu S.
        • de Castro L.F.
        • Jin P.
        • Civin S.
        • Ren J.
        • Reems J.A.
        • et al.
        Manufacturing differences affect human bone marrow stromal cell characteristics and function: Comparison of production methods and products from multiple centers.
        Sci Rep. 2017; 7: 46731
        • Prockop D.J.
        The exciting prospects of new therapies with mesenchymal stromal cells.
        Cytotherapy. 2017; 9: 1-8
        • Boregowda S.V.
        • Krishnappa V.
        • Haga C.L.
        • Ortiz L.A.
        • Phinney D.G.
        A clinical indications prediction scale based on TWIST1 for human mesenchymal stem cells.
        E Bio Medicine. 2016; 4: 62-73
        • Menard C.
        • Pacelli L.
        • Bassi G.
        • Fulong J.
        • Bifari F.
        • Bezler I.
        • et al.
        Clinical-grade mesenchymal stromal cells produced under various good manufacturing practice processes differ in their immunomodulatory properties: standardization of immune quality controls.
        Stem Cells Dev. 2013; 22: 1789-1801
        • Siegel G.
        • Kluba T.
        • Hermanutz-Klein U.
        • Bieback K.
        • Northoff H.
        • Schãfer R.
        Phenotype, donor age and gender affect function of human bone marrow-derived mesenchymal stromal cells.
        BMC Med. 2013; 11: 146
        • Murgia A.
        • Veronesi E.
        • Canini O.
        • Caselli A.
        • D'souza N.
        • Rasini V.
        • et al.
        Potency biomaker signature genes from multiparametric osteogenesis assays: Will cGMP human bone marrow mesenchymal stromal cells make bone?.
        PloS One. 2016; 11e0163629
        • Zaim M.
        • Karaman S.
        • Cetin G.
        • Isik S.
        Donor age and long-term culture affect differentiation and proliferation of human bone marrow mesenchymal stem cells.
        Ann Hematol. 2012; 91: 1175-1186
        • Choi J.R.
        • Yong K.W.
        • Wan Safwani W.K.Z.
        Effect of hypoxia on human adipose-derived mesenchymal stem cells and its potential clinical applications.
        Cell Mol Life Sci. 2017; 74: 2587-2600
        • Kusuma G.D.
        • Carthew J.
        • Lim R.
        • Frith J.E.
        Effect of the microenvironment on mesenchymal stem cell paracrine signaling: Opportunities to engineer the therapeutic effect.
        Stem Cells Dev. 2017; 26: 617-631
        • Maartens J.H.
        • De-Juan-Pardo E.
        • Wunner F.M.
        • Simula A.
        • Voelcker N.H.
        • Barry S.C.
        • et al.
        Challenges and opportunities in the manufacture and expansion of cells for therapy.
        Expert Opin Biol Ther. 2017; 17: 1221-1233
        • Robb K.P.
        • Fitzgerald J.C.
        • Barry F.
        • Viswanathan S.
        Mesenchymal stromal cell therapy: progress in manufacturing and assessments of potency.
        Cytotherapy. 2018; (pii: S1465-3249(18)30675-3[Epub ahead of print])
        • Jossen V.
        • van den Bos C.
        • Eibl R.
        • Eibl D.
        Manufacturing human mesenchymal stem cells at clinical scale: process and regulatory challenges.
        Appl Microbiol Biotechnol. 2018; 102: 3981-3994
        • Trento C.
        • Bernardo M.E.
        • Nagler A.
        • Kuci S.
        • Bornhäuser M.
        • Köhl U.
        • et al.
        Manufacturing mesenchymal stromal cells for the treatment of graft-versus-host disease: A survey among centers affiliated with the european society for blood and marrow transplantation.
        Biol Blood Marrow Transplant. 2018; 24: 2365-2370
        • Kurtzberg J.
        • Prockop S.
        • Teira P.
        • Bittencourt H.
        • Lewis V.
        • Chan K.W.
        • et al.
        Allogeneic human mesenchymal stem cell therapy (remestemcel-L, Prochymal) as a rescue agent for severe refractory acute graft-versus-host disease in pediatric patients.
        Biol Blood Marrow Transplant. 2014; 20: 229-235
        • Panés J.
        • Garcia-Olmo D.
        • Van Assche G.
        • Colombel J.F.
        • Reinisch W.
        • Baumgart D.C.
        • et al.
        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
        • Horn P.
        • Bokermann G.
        • Cholewa D.
        • Bork S.
        • Walenda T.
        • Koch C.
        • et al.
        Impact of individual platelet lysates on isolation and growth of human mesenchymal stromal cells.
        Cytotherapy. 2010; 12: 888-898
        • Schallmoser K.
        • Bartmann C.
        • Rohde E.
        • Reinisch A.
        • Kashofer K.
        • Stadelmeyer E.
        • et al.
        Human platelet lysate can replace fetal bovine serum for clinical-scale expansion of functional mesenchymal stromal cells.
        Transfusion. 2007; 47: 1436-1446
        • Capelli C.
        • Domenghini M.
        • Borleri G.
        • Bellavita P.
        • Poma R.
        • Carobbio A.
        • et al.
        Human platelet lysate allows expansion and clinical grade production of mesenchymal stromal cells from small samples of bone marrow aspirates or marrow filter washouts.
        Bone Marrow Transplant. 2007; 40: 785-791
        • Abdelrazik H.
        • Spaggiari G.M.
        • Chiossone L.
        • Moretta L.
        Mesenchymal stem cells expanded in human platelet lysate display a decreased inhibitory capacity on T- and NK-cell proliferation and function.
        Eur J Immunol. 2011; 41: 3281-3290
        • Oikonomopoulos A.
        • van Deen W.K.
        • Manansala A.T.
        • Lacey P.N.
        • Tomakili T.A.
        • Ziman A.
        • et al.
        Optimization of human mesenchymal stem cell manufacturing: the effects of animal/xeno-free media.
        Sci Rep. 2015; 5: 16570
        • Flemming A.
        • Schallmoser K.
        • Strunk D.
        • Stolk M.
        • Volk H.D.
        • Seifert M.
        Immunomodulative efficacy of bone marrow-derived mesenchymal stem cells cultured in human platelet lysate.
        J Clin Immunol. 2011; 31: 1143-1156
        • Fernandez-Rebollo E.
        • Mentrup B.
        • Ebert R.
        • Franzen J.
        • Abagnale G.
        • Sieben T.
        • et al.
        Human platelet lysate versus fetal calf serum: These supplements do not select for different mesenchymal stromal cells.
        Sci Rep. 2017; 7: 5132
        • Dominici M.
        • Le Blanc K.
        • Mueller I.
        • Slaper-Cortenbach I.
        • Marini F.
        • Krause D.
        • et al.
        Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement.
        Cytotherapy. 2006; 8: 315-317
        • Lin C.S.
        • Xin Z.C.
        • Dai J.
        • Lue T.F.
        Commonly used mesenchymal stem cell markers and tracking labels: Limitations and challenges.
        Histol Histopathol. 2013; 28: 1109-1116
        • Li J.
        • Xu S.Q.
        • Zhao Y.M.
        • Yu S.
        • Ge L.H.
        • Xu B.H.
        Comparison of the biological characteristics of human mesenchymal stem cells derived from exfoliated deciduous teeth, bone marrow, gingival tissue, and umbilical cord.
        Mol Med Rep. 2018; 18: 4969-4977
        • Jeon Y.J.
        • Kim J.
        • Cho J.H.
        • Chung H.M.
        • Chae J.I.
        Comparative analysis of human mesenchymal stem cells derived from bone marrow, placenta, and adipose tissue as sources of cell therapy.
        J Cell Biochem. 2016; 117: 1112-1125
        • Kellner J.
        • Sivajothi S.
        • McNiece I.
        Differential properties of human stromal cells from bone marrow, adipose, liver and cardiac tissues.
        Cytotherapy. 2015; 17: 1514-1523
        • Reinisch A.
        • Etchart N.
        • Thomas D.
        • Hofmann N.A.
        • Fruehwirth M.
        • Sinha S.
        • et al.
        Epigenetic and in vivo comparison of diverse MSC sources reveals an endochondral signature for human hematopoietic niche formation.
        Blood. 2015; 125: 249-260
        • Chan C.K.F.
        • Gulati G.S.
        • Sinha R.
        • Tompkins J.V.
        • Lopez M.
        • Carter A.C.
        • et al.
        Identification of the human skeletal stem cell.
        Cell. 2018; 175: 43-56
        • Sacchetti B.
        • Funari A.
        • Michienzi S.
        • Di Cesare S.
        • Piersanti S.
        • Saggio I.
        • et al.
        Self-renewing osteoprogenitors in bone marrow sinusoids can organize a hematopoietic microenvironment.
        Cell. 2007; 131: 324-336
        • Sorrentino A.
        • Ferracin M.
        • Castelli G.
        • Biffoni M.
        • Tomaselli G.
        • Baiocchi M.
        • et al.
        Isolation and characterization of CD146+ multipotent mesenchymal stromal cells.
        Exp Hematol. 2008; 36: 1035-1046