Abstract
Human bone marrow (BM) derived mesenchymal stem cells (MSC) have high capacity to propagate ex vivo with superior reparative, immunosuppressive, and anti-inflammatory properties. Here we describe standardized protocols and culture conditions that enable the isolation, expansion and maintenance of a highly purified and homogenous population of human MSC. These third party-derived off-the-shelf MSC from healthy human bone marrow donors can potently inhibit mitogenically or allogeneically activated human T cells in proliferation assays. The standard operating procedures described in this chapter can be applied to researchers aiming to enhance MSC immunosuppressive properties and defining MSC mechanisms of action. Importantly, these assays can be incorporated into clinical protocols where the safety and efficacy of human BM MSC can be verified in diseases that are modulated by T cell responses.
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References
Mendez-Ferrer S, Michurina TV, Ferraro F et al (2010) Mesenchymal and haematopoietic stem cells form a unique bone marrow niche. Nature 466:829–834
Friedenstein AJ, Chailakhjan RK, Lalykina KS (1970) The development of fibroblast colonies in monolayer cultures of guinea-pig bone marrow and spleen cells. Cell Tissue Kinet 3:393–403
Dominici M, Le Blanc K, Mueller I et al (2006) Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 8:315–317
Sivanathan KN, Gronthos S, Rojas-Canales D et al (2014) Interferon-gamma modification of mesenchymal stem cells: implications of autologous and allogeneic mesenchymal stem cell therapy in allotransplantation. Stem Cell Rev 10:351–375
Sivanathan KN, Rojas-Canales DM, Hope CM et al (2015) Interleukin-17A-induced human mesenchymal stem cells are superior modulators of immunological function. Stem Cells 33:2850–2863
Sivanathan KN, Rojas-Canales D, Grey ST et al (2017) Transcriptome profiling of IL-17A preactivated mesenchymal stem cells: a comparative study to unmodified and IFN-gamma modified mesenchymal stem cells. Stem Cells Int 2017:1025820
Sivanathan KN, Coates PT (2018) IL-17A-induced mesenchymal stem cells have promising therapeutic value for clinical translation. Kidney Int 93:771–773
Togel FE, Westenfelder C (2012) Kidney protection and regeneration following acute injury: progress through stem cell therapy. Am J Kidney Dis 60:1012–1022
Hickson LJ, Eirin A, Lerman LO (2016) Challenges and opportunities for stem cell therapy in patients with chronic kidney disease. Kidney Int 89:767–778
Casiraghi F, Perico N, Cortinovis M et al (2016) Mesenchymal stromal cells in renal transplantation: opportunities and challenges. Nat Rev Nephrol 12:241–253
Squillaro T, Peluso G, Galderisi U (2016) Clinical trials with mesenchymal stem cells: an update. Cell Transplant 25:829–848
Digirolamo CM, Stokes D, Colter D et al (1999) Propagation and senescence of human marrow stromal cells in culture: a simple colony-forming assay identifies samples with the greatest potential to propagate and differentiate. Br J Haematol 107:275–281
Pochampally R (2008) Colony forming unit assays for MSCs. Methods Mol Biol 449:83–91
Sivanathan KN, Gronthos S, Grey ST et al (2017) Immunodepletion and hypoxia preconditioning of mouse compact bone cells as a novel protocol to isolate highly immunosuppressive mesenchymal stem cells. Stem Cells Dev 26:512–527
Lindemann M (2014) Ex vivo assessment of cellular immune function - applications in patient care and clinical studies. Tissue Antigens 84:439–449
Liu Y, Li YQ, Wang HY et al (2015) Effect of serum choice on replicative senescence in mesenchymal stromal cells. Cytotherapy 17:874–884
Psaltis PJ, Paton S, See F et al (2010) Enrichment for STRO-1 expression enhances the cardiovascular paracrine activity of human bone marrow-derived mesenchymal cell populations. J Cell Physiol 223:530–540
De Andrade AV, Riewaldt J, Wehner R et al (2014) Gamma irradiation preserves immunosuppressive potential and inhibits clonogenic capacity of human bone marrow-derived mesenchymal stromal cells. J Cell Mol Med 18:1184–1193
Acknowledgments
We thank all bone marrow donors and the Royal Adelaide Hospital, Adelaide, South Australia as well as the Australian Red Cross Blood Service for all the buffy coats. We would like to specially thank and acknowledge Svjetlana Kireta and Julie Johnston for their expertise, invaluable input throughout their long-term service in our laboratory at the University of Adelaide and Royal Adelaide Hospital and for proof-reading the manuscript. We thank the Australian Government and the National Health and Medical Council (NHMRC) for K.N.S CJ Martin Biomedical Postdoctoral Early Career Fellowship.
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Sivanathan, K.N., Coates, P.T. (2019). Bone Marrow-Derived Progenitor Cells Mediate Immune Cell Regulation. In: Joglekar, M., Hardikar, A. (eds) Progenitor Cells. Methods in Molecular Biology, vol 2029. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9631-5_17
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DOI: https://doi.org/10.1007/978-1-4939-9631-5_17
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