Skip to main content
SpringerLink
Log in

Scalable Manufacturing of Human Hematopoietic Stem/Progenitor Cells Exploiting a Co-culture Platform with Mesenchymal Stromal Cells

  • Protocol
  • First Online:
Stem Cells and Good Manufacturing Practices

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2286))

  • 932 Accesses

Abstract

In the context of hematopoietic cell transplantation, hematopoietic stem/progenitor cells (HSPC) from the umbilical cord blood (UCB) present several advantages compared to adult sources including higher proliferative capacity, abundant availability and ease of collection, non-risk and painless harvesting procedure, and lower risk of graft-versus-host disease. However, the therapeutic utility of UCB HSPC has been limited to pediatric patients due to the low cell frequency per unit of UCB. The development of efficient and cost-effective strategies to generate large numbers of functional UCB HSPC ex vivo would boost all current and future medical uses of these cells. Herein, we describe a scalable serum-free co-culture system for the expansion of UCB-derived CD34+-enriched cells using microcarrier-immobilized human bone marrow-derived mesenchymal stromal cells as feeder cells.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 99.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 129.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 179.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Kondo M, Wagers AJ, Manz MG, Prohaska SS, Scherer DC, Beilhack GF, Shizuru JA, Weissman IL (2003) Biology of hematopoietic stem cells and progenitors: implications for clinical application. Annu Rev Immunol 21:759–806

    Article  CAS  Google Scholar 

  2. Dahlberg A, Brashem-Stein C, Delaney C, Bernstein ID (2014) Enhanced generation of cord blood hematopoietic stem and progenitor cells by culture with StemRegenin1 and Delta1(Ext-IgG.). Leukemia 28(10):2097–2101

    Article  CAS  Google Scholar 

  3. Ballen KK, Gluckman E, Broxmeyer HE (2013) Umbilical cord blood transplantation: the first 25 years and beyond. Blood 122(4):491–498

    Article  CAS  Google Scholar 

  4. Munoz J, Shah N, Rezvani K, Hosing C, Bollard CM, Oran B, Olson A, Popat U, Molldrem J, McNiece IK, Shpall EJ (2014) Concise review: umbilical cord blood transplantation: past, present, and future. Stem Cells Transl Med 3(12):1435–1443

    Article  Google Scholar 

  5. Broxmeyer HE, Douglas GW, Hangoc G, Cooper S, Bard J, English D, Arny M, Thomas L, Boyse EA (1989) Human umbilical cord blood as a potential source of transplantable hematopoietic stem/progenitor cells. Proc Natl Acad Sci U S A 86(10):3828–3832

    Article  CAS  Google Scholar 

  6. MacMillan ML, Weisdorf DJ, Brunstein CG, Cao Q, DeFor TE, Verneris MR, Blazar BR, Wagner JE (2009) Acute graft-versus-host disease after unrelated donor umbilical cord blood transplantation: analysis of risk factors. Blood 113:2410–2415

    Article  CAS  Google Scholar 

  7. Bear AS, Hanley PJ, Bosque DM, Cruz CR, Kaur I, Liu H, Kebriaei P, Hosing C, Rezvani K, Oran B, De Lima MJ, Bollard CM, Shpall EJ (2014) Low rate of infusional toxicity after expanded cord blood transplantation. Cytotherapy 16(8):1153–1157

    Article  Google Scholar 

  8. Ballen K (2017) Update on umbilical cord blood transplantation. F1000 Res 6:1556

    Article  Google Scholar 

  9. Cohen Y, Nagler A (2004) Umbilical cord blood transplantation—how, when and for whom? Blood Rev 18(3):167–179

    Article  Google Scholar 

  10. Gluckman E (2004) Ex vivo expansion of cord blood cells. Exp Hematol 32(5):410–412

    Article  Google Scholar 

  11. Shpall EJ, Quinones R, Giller R, Zeng C, Baron AE, Jones RB, Bearman SI, Nieto Y, Freed B, Madinger N, Hogan CJ, Slat-Vasquez V, Russell P, Blunk B, Schissel D, Hild E, Malcolm J, Ward W, McNiece IK (2002) Transplantation of ex vivo expanded cord blood. Biol Blood Marrow Transplant 8(7):368–376

    Article  Google Scholar 

  12. Cairo MS, Wagner JE (1997) Placental and/or umbilical cord blood: an alternative source of hematopoietic stem cells for transplantation. Blood 90(12):4665–4678

    Article  CAS  Google Scholar 

  13. Pineault N, Abu-Khader A (2015) Advances in umbilical cord blood stem cell expansion and clinical translation. Exp Hematol 43(7):498–513

    Article  Google Scholar 

  14. Maung KK, Horwitz ME (2019) Current and future perspectives on allogeneic transplantation using ex vivo expansion or manipulation of umbilical cord blood cells. Int J Hematol 110(1):50–58

    Article  Google Scholar 

  15. Kumar S, Geiger H (2017) HSC niche biology and HSC expansion ex vivo. Trends Mol Med 23(9):799–819

    Article  Google Scholar 

  16. Delaney C, Heimfeld S, Brashem-Stein C, Voorhies H, Manger RL, Bernstein ID (2010) Notch-mediated expansion of human cord blood progenitor cells capable of rapid myeloid reconstitution. Nat Med 16(2):232–236

    Article  CAS  Google Scholar 

  17. de Lima M, McNiece I, Robinson SN, Munsell M, Eapen M, Horowitz M, Alousi A, Saliba R, McMannis JD, Kaur I, Kebriaei P, Parmar S, Popat U, Hosing C, Champlin R, Bollard C, Molldrem JJ, Jones RB, Nieto Y, Andersson BS, Shah N, Oran B, Cooper LJ, Worth L, Qazilbash MH, Korbling M, Rondon G, Ciurea S, Bosque D, Maewal I, Simmons PJ, Shpall EJ (2012) Cord-blood engraftment with ex vivo mesenchymal-cell coculture. N Engl J Med 367(24):2305–2315

    Article  Google Scholar 

  18. Zandstra PW, Eaves CJ, Piret JM (1994) Expansion of hematopoietic progenitor cell populations in stirred suspension bioreactors of normal human bone marrow cells. Biotechnology (N Y) 12(9):909–914

    CAS  Google Scholar 

  19. Collins PC, Miller WM, Papoutsakis ET (1998) Stirred culture of peripheral and cord blood hematopoietic cells offers advantages over traditional static systems for clinically relevant applications. Biotechnol Bioeng 59(5):534–543

    Article  CAS  Google Scholar 

  20. Collins PC, Nielsen LK, Patel SD, Papoutsakis ET, Miller WM (1998) Characterization of hematopoietic cell expansion, oxygen uptake, and glycolysis in a controlled, stirred-tank bioreactor system. Biotechnol Prog 14(3):466–472

    Article  CAS  Google Scholar 

  21. Wadhwa PD, Lazarus HM, Koc ON, Jaroscak J, Woo D, Stevens CE, Rubinstein P, Laughlin MJ (2003) Hematopoietic recovery after unrelated umbilical cord-blood allogeneic transplantation in adults treated with in vivo stem cell factor (R-MetHuSCF) and filgrastim administration. Leuk Res 27(3):215–220

    Article  CAS  Google Scholar 

  22. Kedong S, Xiubo F, Tianqing L, Macedo HM, LiLi J, Meiyun F, Fangxin S, Xuehu M, Zhanfeng C (2010) Simultaneous expansion and harvest of hematopoietic stem cells and mesenchymal stem cells derived from umbilical cord blood. J Mater Sci Mater Med 21(12):3183–3193

    Article  Google Scholar 

  23. Andrade-Zaldivar H, Kalixto-Sanchez MA, de la Rosa AP, De Leon-Rodriguez A (2011) Expansion of human hematopoietic cells from umbilical cord blood using roller bottles in CO2 and CO2-free atmosphere. Stem Cells Dev 20(4):593–598

    Article  Google Scholar 

  24. Schmelzer E, Finoli A, Nettleship I, Gerlach JC (2015) Long-term three-dimensional perfusion culture of human adult bone marrow mononuclear cells in bioreactors. Biotechnol Bioeng 112(4):801–810

    Article  CAS  Google Scholar 

  25. Xue C, Kwek KY, Chan JK, Chen Q, Lim M (2014) The hollow fiber bioreactor as a stroma-supported, serum-free ex vivo expansion platform for human umbilical cord blood cells. Biotechnol J 9(7):980–989

    Article  CAS  Google Scholar 

  26. Andrade PZ, da Silva CL, dos Santos F, Almeida-Porada G, Cabral JM (2011) Initial CD34+ cell-enrichment of cord blood determines hematopoietic stem/progenitor cell yield upon ex vivo expansion. J Cell Biochem 112(7):1822–1831

    Article  CAS  Google Scholar 

  27. Andrade PZ, dos Santos F, Almeida-Porada G, da Silva CL, S Cabral JM (2010) Systematic delineation of optimal cytokine concentrations to expand hematopoietic stem/progenitor cells in co-culture with mesenchymal stem cells. Mol Biosyst 6(7):1207–1215

    Article  CAS  Google Scholar 

  28. da Silva CL, Goncalves R, Crapnell KB, Cabral JM, Zanjani ED, Almeida-Porada G (2005) A human stromal-based serum-free culture system supports the ex vivo expansion/maintenance of bone marrow and cord blood hematopoietic stem/progenitor cells. Exp Hematol 33(7):828–835

    Article  Google Scholar 

  29. Andrade PZ, de Soure AM, Dos Santos F, Paiva A, Cabral JM, da Silva CL (2015) Ex vivo expansion of cord blood haematopoietic stem/progenitor cells under physiological oxygen tensions: clear-cut effects on cell proliferation, differentiation and metabolism. J Tissue Eng Regen Med 9(10):1172–1181

    Article  CAS  Google Scholar 

  30. Dos Santos F, Andrade PZ, Abecasis MM, Gimble JM, Chase LG, Campbell AM, Boucher S, Vemuri MC, Silva CL, Cabral JM (2011) Toward a clinical-grade expansion of mesenchymal stem cells from human sources: a microcarrier-based culture system under xeno-free conditions. Tissue Eng Part C Methods 17(12):1201–1210

    Article  Google Scholar 

  31. Moore MA, Hoskins I (1994) Ex vivo expansion of cord blood-derived stem cells and progenitors. Blood Cells 20(2–3):468–479. discussion 479-81

    CAS  PubMed  Google Scholar 

  32. Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, Deans R, Keating A, Prockop D, Horwitz E (2006) Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 8(4):315–317

    Article  CAS  Google Scholar 

  33. de Soure AM, Fernandes-Platzgummer A, Moreira F, Lilaia C, Liu SH, Ku CP, Huang YF, Milligan W, Cabral JMS, da Silva CL (2017) Integrated culture platform based on a human platelet lysate supplement for the isolation and scalable manufacturing of umbilical cord matrix-derived mesenchymal stem/stromal cells. J Tissue Eng Regen Med 11(5):1630–1640

    Article  Google Scholar 

Download references

Acknowledgments

Funding is received by Institute for Bioengineering and Biosciences (iBB) from Portuguese Funding for Science and Technology (UIDB/04565/2020) and from Programa Operacional Regional (POR) de Lisboa 2020 through the project PRECISE—Accelerating progress toward the new era of precision medicine (Project N. 16394). Current affiliation of Pedro Z. Andrade is Biosurfit SA., Azambuja, Portugal.

Author information

Authors and Affiliations

  1. Department of Bioengineering and iBB-Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal

    Ana Fernandes-Platzgummer, Pedro Z. Andrade, Joaquim M. S. Cabral & Cláudia Lobato da Silva

Authors
  1. Ana Fernandes-Platzgummer
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Pedro Z. Andrade
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Joaquim M. S. Cabral
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Cláudia Lobato da Silva
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Cláudia Lobato da Silva .

Editor information

Editors and Affiliations

  1. Ottawa, ON, Canada

    Kursad Turksen

Rights and permissions

Reprints and permissions

Copyright information

© 2020 Springer Science+Business Media New York

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Fernandes-Platzgummer, A., Andrade, P.Z., Cabral, J.M.S., da Silva, C.L. (2020). Scalable Manufacturing of Human Hematopoietic Stem/Progenitor Cells Exploiting a Co-culture Platform with Mesenchymal Stromal Cells. In: Turksen, K. (eds) Stem Cells and Good Manufacturing Practices. Methods in Molecular Biology, vol 2286. Humana, New York, NY. https://doi.org/10.1007/7651_2020_289

Download citation

  • DOI: https://doi.org/10.1007/7651_2020_289

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1326-9

  • Online ISBN: 978-1-0716-1327-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation