, Volume 56, Issue 3, pp 151–160 | Cite as

Expansion of human hematopoietic stem cells for transplantation: trends and perspectives

  • Hera Andrade-Zaldívar
  • Leticia Santos
  • Antonio De León RodríguezEmail author


Umbilical cord blood transplantation is clinically limited by its low progenitor cell content. Ex vivo expansion has become an alternative to increase the cell dose available for transplants. Expansion has been evaluated in several ways such as static cultures combining growth factors or mimicking the natural microenvironment using co-culture systems. However, static cultures have a small volume capacity and therefore large-scale expansion has been addressed using bioreactors. These and other biotechnological approaches for the expansion of hematopoietic progenitors and their utility to study several aspects of hematopoietic stem cell biology are discussed here.


2D-culture 3D-culture Human cells Leukemia Rotating wall vessel Stirred tank Transplant 



This work was partially supported by CONACYT-Fondos Sectoriales Salud Grant 13993 and Grant I0010-2007-01, the sabbatical fellowship 79973 and PhD scholarship 103403.


  1. Astori G, Adami V, Mambrini G, Bigi L, Cilli M, Facchini A, Falsaca E, Malangone W (2005) Evaluation of ex vivo expansion and engraftment in NOD/SCID mice of umbilical cord blood CD34+ cells using the DIDECO “Pluricell System”. Bone Marrow Transplant 35:1101–1106CrossRefGoogle Scholar
  2. Attema JL, Papathanasiou P, Forsberg EC, Xu J, Smale ST, Weissman IL (2007) Epigenetic characterization of hematopoietic stem cell differentiation using miniChIP and bisulfite sequencing analysis. Proc Natl Acad Sci USA 104:12371–12376CrossRefGoogle Scholar
  3. Bornstein R, Flores AI, Montalban MA, del Rey MJ, de la Serna J, Gilsanz F (2005) A modified cord blood collection method achieves sufficient levels for transplantation in most adult patients. Stem cells 23:324–334Google Scholar
  4. Cabrita GJ, Fereira BS, da Silva CL, Gonçalves R, Almeida-Porada G, Cabral JM (2003) Hematopoietic stem cells: from the bone to the bioreactor. Trends Biotechnol 21:233–240CrossRefGoogle Scholar
  5. Chivu M, Diaconu CC, Bleotu C, Alexiu I, Brasoveanu L, Cernescu C (2004) The comparison of different protocols for expansion of umbilical-cord blood hematopoietic stem cells. J Cell Mol Med 8:223–231CrossRefGoogle Scholar
  6. 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 14:466–472CrossRefGoogle Scholar
  7. De Leon A, Mayani H, Ramirez O (1998) Design, characterization and application of a minibioreactor for the culture of human hematopoietic cells under controlled conditions. Cytotechnology 28(1–3):127–138CrossRefGoogle Scholar
  8. De León A, Barba-de la Rosa AP, Mayani H, Galindo E, Ramírez OT (2001) Two useful dimensionless parameters that combine physiological, operational and bioreactor design parameters for improved control of dissolved oxygen. Biotechnol Lett 23:1051–1056CrossRefGoogle Scholar
  9. Devine SM, Lazarus HM, Emerson SG (2003) Clinical application of hematopoietic progenitor cell expansion: current status and future prospects. Bone Marrow Transplant 31:241–252CrossRefGoogle Scholar
  10. Dykstra B, de Hann G (2008) Hematopoietic stem cell aging and self-renewal. Cell Tissue Res 331:91–101CrossRefGoogle Scholar
  11. Ehring B, Biber TM, Upton TM, Plosky D, Pykett M, Rosenzweig M (2003) Expansion of HPCs from cord blood in a novel 3D matrix. Cytotherapy 5:490–499CrossRefGoogle Scholar
  12. Feng Q, Chai C, Jiang XS, Leong KW, Mao HQ (2006) Expansion of engrafting human hematopoietic stem/progenitor cells in three-dimensional scaffolds with surface-immobilized fibronectin. J Biomed Mater Res 78:781–791CrossRefGoogle Scholar
  13. Fujimoto N, Fujita S, Takashi T, Toguchida J, Kenji I, Hiroshi S, Hiroo I (2007) Microencapsulated an feeder cells as a source of soluble factors for expansion of CD34+ hematopoietic stem cells. Biomaterials 28:4795–4805CrossRefGoogle Scholar
  14. Gerecht-Nir S, Cohen S, Itskovitz-Eldor J (2004) Bioreactor cultivation enhances the efficiency of human embryoid body (hEB) formation and differentiation. Biotechnol Bioeng 86:493–502CrossRefGoogle Scholar
  15. Gluckman E, Broxmeyer HE, Auerbach AD (1989) Hematopoietic reconstitution of a patient with Fanconi anemia by means of umbilical cord blood from an HLA-identical sibling. N Engl J Med 321:1174–1178CrossRefGoogle Scholar
  16. Ho L, Greene CL, Schmidt AW, Huang LH (2004) Cultivation of HEK 293 cell line and production of a member of the superfamily of G-protein coupled receptors for drug discovery applications using a highly efficient novel bioreactor. Cytotechnology 45:117–123CrossRefGoogle Scholar
  17. Hofmeister CC, Zhang J, Knight KL, Le P, Stiff PJ (2007) Ex vivo expansion of umbilical cord blood stem cells for transplantation: growing knowledge from the hematopoietic niche. Bone Marrow Transplant 39:11–23CrossRefGoogle Scholar
  18. Introna M, Franceschetti M, Ciocca A, Borleri G, Conti E, Golay J, Rambaldi A (2006) Rapid and massive expansion of cord blood-derived cytokine-induced killer cells: an innovative proposal for the treatment of leukemia relapse after cord blood transplantation. Bone Marrow Transplant 38:621–627CrossRefGoogle Scholar
  19. Jang YK, Jung DH, Jung MH, Kim DH, Yoo KH, Sung KW, Koo HH, Oh W, Yang YS, Yang SE (2006) Mesenchymal stem cells feeder layer from human umbilical cord blood for ex vivo expanded growth and proliferation of hematopoietic progenitor cells. Ann Hematol 85:212–225CrossRefGoogle Scholar
  20. Josephson R, Sykes G, Liu Y, Ording C, Xu W, Zeng X, Shin S, Loring J, Maitra A, Rao M, Auerbach J (2006) A molecular scheme for improved characterization of human embryonic stem cell lines. BMC Biol 4:28CrossRefGoogle Scholar
  21. Kasper C, Suck K, Anton F, Scheper T, Kall S, van Griensven M (2007) A newly developed rotating bed bioreactor for bone tissue engineering. In: Ashammakhi N, Reis R, Chiellini E (eds) Topics in tissue engineering. Oulu, Finland, pp 1–15Google Scholar
  22. Koller MR, Palsson MA, Manchel I,Maher RJ, Palsson BO (1998) Tissue culture surface characteristics influence the expansion of human bone marrow cells. Biomaterials 19:1963–1972CrossRefGoogle Scholar
  23. Kwon J, Kim BS, Kim MJ, Park HW (2003) Suspension culture of hematopoietic stem cells in stirred bioreactors. Biotechnol Lett 25:179–182CrossRefGoogle Scholar
  24. Lim M, Ye H, Panoskaltsis N, Drakakis EM, Yue X, Cass AE, Radomska A, Mantalaris A (2007) Intelligent bioprocessing for haemotopoietic cell cultures using monitoring and design of experiments. Biotechnol Adv 25:353–368CrossRefGoogle Scholar
  25. Liu Y, Liu T, Fan X, Ma X, Cui Z (2006) Ex vivo expansion of hematopoietic stem cells derived from umbilical cord blood in rotating wall vessel. J Biotechnol 124:592–601CrossRefGoogle Scholar
  26. Madlambayan GJ, Rogers I, Purpura KA, Ito C, Yu M, Kirouac D, Casper RF, Zandstra PW (2006) Clinically relevant expansion of hematopoietic stem cells with conserved function in a single-use, closed-system bioprocess. Biol Blood Marrow Transplant 12:1020–1030CrossRefGoogle Scholar
  27. Martin Y, Vermette P (2005) Bioreactors for tissue mass culture: design, characterization, and recent advances. Biomaterials 26:7481–7503CrossRefGoogle Scholar
  28. Matsunaga T, Tanaka I, Kobune M, Kawano Y, Tanaka M, Kuribayashi K, Iyama S, Sato T, Sato Y, Takimoto R, Takayama T, Kato J, Ninomiya T, Hamada H, Niitsu Y (2006) Ex vivo large-scale generation of human platelets from cord blood CD34+ cells. Stem Cells 24:2877–2887CrossRefGoogle Scholar
  29. Mohamed AA, Ibrahim AM, El-Masry MW, Manssur IM, Khroshied MA, Gouda HM, Riad RM (2006) Ex vivo expansion of stem cells: defining optimum conditions using various cytokines. Lab Hematol 12:86–93CrossRefGoogle Scholar
  30. Muller-Sieburg CE, Sieburg HB (2006) The GOD of hematopoietic stem cells, a clonal diversity model of the stem cell compartment. Cell Cycle 5:394–398Google Scholar
  31. Ohi S, Roach AN, Ramsahai S, Kim BC, Fitzgerald W, Riley DA, Gonda SR (2004) The hematopoietic stem cell therapy for exploration of deep space. New Frontiers & Future Concepts. AIP Conf Proc 699:938–950CrossRefGoogle Scholar
  32. Piacibello W, Sanavio F, Garetto L, Severino A, Dane A, Gammaitoni L, Aglietta M (1998) Differential growth factor requirement of primitive cord blood hematopoietic stem cell for self-renewal and amplification vs proliferation and differentiation. Leukemia 12:718–727CrossRefGoogle Scholar
  33. Qiu H, Fujimori Y, Kai S, Fujibayashi Y, Nishioka K, Hara H (2003) Establishment of mouse embryonic fibroblast cell lines that promote ex vivo expansion of human cord blood CD34+ hematopoietic progenitors. J Hematother Stem Cell Res 12:39–46CrossRefGoogle Scholar
  34. Qunliang L, Qiwei L, Haibo C, Wen-Song T (2006) A comparative gene-expression analysis of CD34+ hematopoietic stem and progenitor cells grown in static and stirred culture systems. Cell Mol Biol Lett 11:475–487CrossRefGoogle Scholar
  35. Robinson S, Niu T, de Lima M, Ng J, Yang H, McManis J, Karandish S, Sadeghi T, Fu P, del Angel M, O’Connor S, Dhamplin R, Shpall E (2005) Ex vivo expansion of umbilical cord blood. Cytotherapy 7:243–250CrossRefGoogle Scholar
  36. Saulnier N, Di Campli C, Zocco MA, Di Gioacchino G, Novi M, Gasbarrini A (2005) From stem cell to solid organ. Bone marrow, peripheral blood or umbilical cord blood as favorable source? Eur Rev Med Pharmacol Sci 9:315–324Google Scholar
  37. Schoemans H, Theunissen K, Maertens J, Boogaerts M, Verfaillie C, Wagner J (2006) Adult umbilical cord blood transplantation: a comprehensive review. Bone Marrow Transplant 38:83–93CrossRefGoogle Scholar
  38. Schubert H, Garrn I, Berthold A, Knauff WU, Reufi B, Fietz T, Gross UM (2004) Culture of haematopoietic cells in a 3D bioreactor made of Al2O3 or apatite foam. J Mater Sci Mater Med 4:331–334CrossRefGoogle Scholar
  39. Tong J, Gianni AM, Siena S, Srour EF, Bregni M, Hoffman R (1994) Primitive hematopoietic progenitor cells are present in peripheral blood autografts. Blood Cells 20:351–362, discussion 362–363Google Scholar
  40. Unsworth BR, Lelkes PI (1998) Growing tissues in microgravity. Nat Med 4:901–907CrossRefGoogle Scholar
  41. Van Zant G, Rummel SA, Koller MR, Larson DB, Drubachevsky I, Palsson M, Emerson SG (1994) Expansion in bioreactors of human progenitor populations from cord blood and mobilized peripheral blood. Blood Cells 20:482–491Google Scholar
  42. Xie C, Jia B, Xiang Y, Wang L, Wang G, Huang G, McNiece IK, Wang J. (2006) Support of hMSCs transduced with TPO/FL genes to expansion of umbilical cord CD34+ cells in indirect co-culture. Cell Tissue Res 326:101–110CrossRefGoogle Scholar
  43. Yamaguchi M, Hirayama F, Murahashi H, Azuma H, Sato N, Miyakazu H, Fukazawa K, Sawada K, Koike T, Kuwabara M, Ikeda H, Ikebuchi K (2002) Ex vivo expansion of UC blood primitive hematopoietic progenitors and transplantable stem cells using human primary BM stromal cells and human AB serum. Cytotherapy 4:109–118CrossRefGoogle Scholar
  44. Yang S, Cai H, Jin H, Tan WS (2008) Hematopoietic reconstitution of CD34+ cells grown in static and stirred culture systems in NOD/SCID mice. Biotechnol Lett 30:61–65CrossRefGoogle Scholar
  45. Yao CL, Fena SH, Lin XZ, Chu IM, Hsieh TB, Hwang SM (2006) Characterization of serum-free ex vivo-expanded hematopoietic cells derived from human umbilical cord blood CD133+ cells. Stem Cells Dev 15:70–78CrossRefGoogle Scholar
  46. Yoo ES, Lee KE, Seo JW, Yoo EH, Lee MA, Im SA, Mun YC, Lee SN, Huh JW, Kim MJ, Jo DY, Ahn JY, Lee SM, Chung WS, Kim JH, Seong CM (2003) Adherent cells generated during long-term culture of human cord blood CD34+ cells have characteristics of endothelial cells and beneficial effect on cord blood ex vivo expansion. Stem Cells 21:228–235CrossRefGoogle Scholar
  47. Zhai QL, Qiu LG, Li Q, Meng HX, Han JL, Herzig RH, Han ZC (2004) Short-term ex vivo expansion sustains the homing-related properties of umbilical cord blood hematopoietic stem and progenitor cells. Haematologica 89:265–272Google Scholar
  48. Zhang Y, Chai C, Jiang XS, Teoh SH, Leong KW (2006) Co-culture of umbilical cord blood CD34+ cells with human mesenchymal stem cells. Tissue Eng 12:2161–2170CrossRefGoogle Scholar
  49. Zubler RH (2006) Ex vivo expansion of hematopoietic stem cells and gene therapy development. Swiss Med Wkly 136:795–799Google Scholar

Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Hera Andrade-Zaldívar
    • 1
  • Leticia Santos
    • 1
  • Antonio De León Rodríguez
    • 1
    Email author
  1. 1.División de Biología MolecularInstituto Potosino de Investigación Científica y TecnológicaSan Luis PotosiMexico

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