Advertisement

Isolation and Manufacture of Clinical-Grade Bone Marrow-Derived Human Mesenchymal Stromal Cells

  • Renuka P. Miller
  • Patrick J. HanleyEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1416)

Abstract

Mesenchymal stromal cells (MSCs) are multipotent cells with both regenerative and immunomodulatory capacities. These unique properties make them appealing as a biologic, with multiple phase 1–3 clinical trials currently testing their safety and efficacy. Although expanding MSCs does not require extensive manipulation, expanding MSCs for use in clinical trials does require the knowledge and safety that are delineated in current good manufacturing practices (GMPs). Here we briefly detail the characteristics of MSCs and considerations for expanding them for clinical use. We then include a step-by-step protocol for expanding MSCs for early phase clinical trials, with important notes to consider during the expansion of these MSCs.

Key words

Mesenchymal stromal cells Clinical grade Good manufacturing practices Cell therapy 

Notes

Acknowledgments

The authors would like to thank our collaborators at the Baylor College of Medicine, namely, Dr. Adrian Gee, Dr. Zhuyong Mei, and Dr. Helen Heslop. We would also like to thank the Program for Cell Enhancement and Technologies for Immunotherapy (CETI) director, Dr. Catherine Bollard, for her support and guidance.

References

  1. 1.
    Friedenstein AJ, Deriglasova UF, Kulagina NN et al (1974) Precursors for fibroblasts in different populations of hematopoietic cells as detected by the in vitro colony assay method. Exp Hematol 2:83–92PubMedGoogle Scholar
  2. 2.
    Hanley PJ, Mei Z, da Graca Cabreira-Hansen M et al (2013) Manufacturing mesenchymal stromal cells for phase I clinical trials. Cytotherapy 15:416–422CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Zuk PA, Zhu M, Ashjian P et al (2002) Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell 13: 4279–4295CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Ryan JM, Barry FP, Murphy JM et al (2005) Mesenchymal stem cells avoid allogeneic rejection. J Inflamm (Lond) 2:8CrossRefGoogle Scholar
  5. 5.
    Majumdar M, Keane-Moore M, Buyaner D et al (2003) Characterization and functionality of cell surface molecules on human mesenchymal stem cells. J Biomed Sci 10:228–241CrossRefPubMedGoogle Scholar
  6. 6.
    Tse WT, Pendleton JD, Beyer WM et al (2003) Suppression of allogeneic T-cell proliferation by human marrow stromal cells: implications in transplantation. Transplantation 75:389–397CrossRefPubMedGoogle Scholar
  7. 7.
    Li YP, Paczesny S, Lauret E et al (2008) Human mesenchymal stem cells license adult CD34+ hemopoietic progenitor cells to differentiate into regulatory dendritic cells through activation of the Notch pathway. J Immunol 180:1598–1608CrossRefPubMedGoogle Scholar
  8. 8.
    Abumaree M, Al Jumah M, Pace RA et al (2012) Immunosuppressive properties of mesenchymal stem cells. Stem Cell Rev 8:375–392CrossRefPubMedGoogle Scholar
  9. 9.
    Le Blanc K, Rasmusson I, Sundberg B et al (2004) Treatment of severe acute graft-versus-host disease with third party haploidentical mesenchymal stem cells. Lancet 363:1439–1441CrossRefPubMedGoogle Scholar
  10. 10.
    Yin F, Battiwalla M, Ito S et al (2014) Bone marrow mesenchymal stromal cells to treat tissue damage in allogeneic stem cell transplant recipients: correlation of biological markers with clinical responses. Stem Cells 32:1278–1288CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Lazarus HM, Koc ON, Devine SM et al (2005) Cotransplantation of HLA-identical sibling culture-expanded mesenchymal stem cells and hematopoietic stem cells in hematologic malignancy patients. Biol Blood Marrow Transplant 11:389–398CrossRefPubMedGoogle Scholar
  12. 12.
    Forbes GM, Sturm MJ, Leong RW et al (2014) A phase 2 study of allogeneic mesenchymal stromal cells for luminal Crohn’s disease refractory to biologic therapy. Clin Gastroenterol Hepatol 12:64–71CrossRefPubMedGoogle Scholar
  13. 13.
    Horwitz EM, Gordon PL, Koo WK et al (2002) Isolated allogeneic bone marrow-derived mesenchymal cells engraft and stimulate growth in children with osteogenesis imperfecta: Implications for cell therapy of bone. Proc Natl Acad Sci 99:8932–8937CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Wu Y, Chen L, Scott PG et al (2007) Mesenchymal stem cells enhance wound healing through differentiation and angiogenesis. Stem Cells 25:2648–2659CrossRefPubMedGoogle Scholar
  15. 15.
    Baxter MA, Wynn RF, Jowitt SN et al (2004) Study of telomere length reveals rapid aging of human marrow stromal cells following in vitro expansion. Stem Cells 22:675–682CrossRefPubMedGoogle Scholar
  16. 16.
    Stolzing A, Jones E, McGonagle D et al (2008) Age-related changes in human bone marrow-derived mesenchymal stem cells: consequences for cell therapies. Mech Ageing Dev 129:163–173CrossRefPubMedGoogle Scholar
  17. 17.
    Kern S, Eichler H, Stoeve J et al (2006) Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue. Stem Cells 24:1294–1301CrossRefPubMedGoogle Scholar
  18. 18.
    Hanley PJ (2014) Finessing the manufacture of mesenchymal stromal cells. Cytotherapy 16:711–712CrossRefPubMedGoogle Scholar
  19. 19.
    Hanley PJ, Mei Z, Durett AG et al (2014) Efficient manufacturing of therapeutic mesenchymal stromal cells with the use of the Quantum Cell Expansion System. Cytotherapy 16:1048–1058CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Timmins NE, Kiel M, Günther M et al (2012) Closed system isolation and scalable expansion of human placental mesenchymal stem cells. Biotechnol Bioeng 109:1817–1826CrossRefPubMedGoogle Scholar
  21. 21.
    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 8:315–317CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Program for Cell Enhancement and Technologies for ImmunotherapyCenter for Cancer and Immunology ResearchWashingtonUSA
  2. 2.Children’s National Health SystemWashingtonUSA
  3. 3.Division of Blood and Marrow TransplantationWashingtonUSA
  4. 4.Sheikh Zayed Institute for Pediatric Surgical InnovationWashingtonUSA
  5. 5.The George Washington UniversityWashingtonUSA

Personalised recommendations