Haematopoietic Culture Systems

  • L. Safinia
  • N. Panoskaltsis
  • A. Mantalaris


Haematopoietic stem cells have been applied successfully in the clinic for over 30 years. This experience, the relative ease with which HSCs can be identified and obtained from a variety of sources and the potential plasticity of these cells makes them ideal for use in haematologic and non-haematologic conditions. Despite these advantages and the significant progress that has been made in the characterisation of factors that govern haematopoiesis, enrichment and ex vivo expansion of repopulating (and possibly plastic) HSCs remains elusive. Conventional 2-D cultures are insufficient to meet the complex demands required and small deviations in the culture parameters can profoundly affect the final cell output. The application of factorial and composite designs to HSC cultures is required in order to fully appreciate the effects and interdependence of stimulatory and inhibitory factors as well as the culture parameters on haematopoietic culture systems. Furthermore, ex vivo expanded HSCs must be safe to use in humans and meet regulations guided by good manufacturing practice (GMP) requirements for clinical therapeutics which includes the development of suitable, closed culture systems that can be easily controlled and monitored. The engineering of optimal haematopoietic cell culture systems requires the design of new expansion systems that mimic the in vivo bone marrow environment that is able to self-regulate and operate under reliable and reproducible conditions. Such a system would offer a broad spectrum of possibilities for different culture strategies in the cultivation of various cell types — from stem cells to differentiated cells for gene, cellular, and tissue therapies.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Abboud CN, Lichtman MA. 2001. Structure of the Marrow and the Hemopoietic Microenvironment. In: Seligsohn U, editor. Hematology. New York: McGraw-Hill. p 29–58.Google Scholar
  2. Aiuti A, Friedrich C, Sieff C, Gutierrez-Ramos J. 1998. Identification of distinct elements of the stromal microenvironment that control human hematopoietic stem/progenitor cell growth and differentiation. Experimental Hematology 26:143–157.PubMedGoogle Scholar
  3. Akatov V, Lezhnev, EI., Vexler, AM., Kublik, LN. 1985. Low pH value of pericellular medium as a factor limiting cell proliferation in dense cultures. Experimental Cell Research 160:412–418.CrossRefPubMedGoogle Scholar
  4. Alley C, MacDermott, RP., Stewart, CC. 1983. The Effect of Serum on Human Marrow Mononuclear Cell Proliferation and Maturation. Journal of the Reticuloendothelial Society 34:271–278.PubMedGoogle Scholar
  5. Almeida-Porada G, Brown, RL., MacKintosh, FR., Zanjani, ED. 2000. Evaluation of Serum-Free Culture Conditions Able to Support the Ex Vivo Expansion and Engraftment of Human Hematopoietic Stem CElls in Human-to Sheep Xenograft Model. Journal of Hematotherapy & Stem Cell Research 9:683–693.CrossRefGoogle Scholar
  6. Audet J, Zanstra, PW., Eaves, CJ., Piret, JM. 1998. Advances in hematopoietic stem cell culture. Current Opinion in Biotechnology 9:146–151.CrossRefPubMedGoogle Scholar
  7. Bachier CR, Gokmen E, Teale J, Lanzkron S, Childs C, Franklin W, Shpall E, Douville J, Weber S, Muller T and others. 1999. Ex-vivo expansion of bone marrow progenitor cells for hematopoietic reconstitution following high-dose chemotherapy for breast cancer. Experimental Hematology 27:615–623.CrossRefPubMedGoogle Scholar
  8. Bagley J, Rosenzweig, M., Mark, DF., Pykett, MJ. 1999. Extended culture of multipotent hematopoietic progenitors without cytokine augmentation in a novel three-dimensional device. Experimental Hematology 27:496–504.CrossRefPubMedGoogle Scholar
  9. Barnett MJ, Eaves CJ, Phillips GL, Gascoyne RD, Hogge DE, Horsman DE, Humphries RK, Klingemann HG, Lansdorp PM, Nantel SH and others. 1994. Autografting with cultured marrow in chronic myeloid leukemia: results of a pilot study. Blood 84:724–732.PubMedGoogle Scholar
  10. Bhatia M, Bonnet D, Wu D, Murdoch B, Wrana J, Gallacher L, Dick JE. 1999. Bone morphogenetic proteins regulate the developmental program of human hematopoietic stem cells. Journal of Experimental Medicine 189:1139–1148.CrossRefPubMedGoogle Scholar
  11. Bhatia M, McGlave PB, Dewald GW, Blazar BR, Verfaillie CM. 1995. Abnormal function of the bone marrow microenvironment in chronic myelogenous leukemia: role of malignant stromal macrophages. Blood 85:3636–3645.PubMedGoogle Scholar
  12. Bohmer R. 1989. Interaction of Serum and Colony-Stimulating Factor for Survival of a Factor-Dependent Hemopoietic Progenitor Cell Line. Journal of Cellular Physiology 139:531–537.CrossRefPubMedGoogle Scholar
  13. Brandt J, Bhalla, K., Hoffman, R. 1994. Effects of interleukin-3 and c-kit ligand on the survival of various classes of human hematopoietic progenitor cells. Blood 83(6):1507–1514.PubMedGoogle Scholar
  14. Bregni M, Magni M, Siena S, Di Nicola M, Bonadonna G, Gianni AM. 1992. Human peripheral blood hematopoietic progenitors are optimal targets of retroviral-mediated gene transfer. Blood 80:1418–1422.PubMedGoogle Scholar
  15. Broudy VC. 1997. Stem cell factor and hematopoiesis. Blood 90:1354–1364.Google Scholar
  16. Brown R, Xu, FS., Dusing, SK., Li, Q., Fischer, R., Patchen, M. 1997. Serum-Free Conditions for Cells Capable of Producing Long-Term Survival in Lethally Irradiated Mice. Stem Cells 15:237–245.PubMedGoogle Scholar
  17. Broxmeyer HE, Sherry B, Cooper S, Lu L, Maze R, Beckmann MP, Cerami A, Ralph P. 1993. Comparative analysis of the human macrophage inflammatory protein family of cytokines (chemokines) on proliferation of human myeloid progenitor cells. Interacting effects involving suppression, synergistic suppression, and blocking of suppression. Journal of Immunology 150:3448–3458.Google Scholar
  18. Brugger W, Mocklin W, Heimfeld S, Berenson RJ, Mertelsmann R, Kanz L. 1993. Ex vivo expansion of enriched peripheral blood CD34+ progenitor cells by stem cell factor, interleukin-1 (IL-1), IL-6, IL-3, interferon g and erythropoietin. Blood 81:2579–2584.PubMedGoogle Scholar
  19. Cabral JMS. 2001. Ex vivo expansion of hematopoietic stem cells in bioreactors. Biotechnology Letters 23:741–751.CrossRefGoogle Scholar
  20. Cairo MS, Wagner JE. 1997. Placental and/or umbilical cord blood: an alternative source of hematopoietic stem cells for transplantation. Blood 90:4664–4671.Google Scholar
  21. Caldwell J, Locey B, Clarke MF, Emerson SG, Palsson BO. 1991a. Influence of medium exchange schedules on metabolic, growth, and GM-CSF secretion rates of genetically engineered NIH-3T3 cells. Biotechnology Progress 7:1–8.CrossRefPubMedGoogle Scholar
  22. Caldwell J, Palsson BO, Locey B, Emerson SG. 1991b. Culture perfusion schedules influence the metabolic activity and granulocyte-macrophage colony-stimulating factor production rates of human bone marrow stromal cells. Journal of Cellural Physiology 147:344–353.CrossRefGoogle Scholar
  23. Carswell K, Papoutsakis, ET. 2000. Extracellular pH affects the proliferation of cultured human T-cells and their expression of the interleukin-2 receptor. Journal of Immunology 23(6):669–674.Google Scholar
  24. Chabannon C, Olivero S, Blaise D, Maraninchi D, Viens P. 2000. Ex vivo expansion of human hematopoietic progenitors and cells to support high-dose chemoradiation therapy: Five years of clinical experience. Cytokines, Cellular & Molecular Therapy 6:97–108.CrossRefGoogle Scholar
  25. Charbord P. 2001. Microenvironmental Cell Populations Essential for the Support of Hematopoietic Stem Cells. In: Zon LI, editor. Hematopoiesis: A Developmental Approach. New York: Oxford University Press. p 691–701.Google Scholar
  26. Chute JP, Saini AA, Kampen RL, Wells MR, Davis TA. 1999. A comparative study of the cell cycle status and primitive cell adhesion molecule profile of human CD34+ cells cultured in stroma-free versus porcine microvascular endothelial cell cultures. Experimental Hematology 27:370–379.CrossRefPubMedGoogle Scholar
  27. Cipolleschi M, D’Ippolito, G., Bernabei, PA., Caporale, R., Nannini, R., Mariani, M., Fabbiani, M., Rossi-Ferrini, P., Olivotto, M., Sbarba, PD. 1997. Severe hypoxia enhances the formation of erythroid bursts from human cord blood cells and the maintenance of BFU-E in vitro. Experimental Hematology 25:1187–1194.PubMedGoogle Scholar
  28. Cipolleschi MG, Dello Sbarba P, Olivotto M. 1993. The role of hypoxia in the mainetance of hematopoietic stem cells. Blood 82:2031–2037.PubMedGoogle Scholar
  29. Civin CI, Almeida-Porada G, Lee MJ, Olweus J, Terstappen LW, Zanjani ED. 1996. Sustained, retransplantable, multilineage engraftment of highly purified adult human bone marrow stem cells in vivo. Blood 88:4102–4109.PubMedGoogle Scholar
  30. Collins P, Miller, WM., Papoutsakis, ET. 1996. Ex vivo culture systems for hematopoietic cells. Current Opinion in Biotechnology 7(2):223–230.CrossRefPubMedGoogle Scholar
  31. Collins PC, Miller ME, Papoutsakis ET. 1998. Stirred culture of peripheral and cord blood hematopoietic cells offers advantages over traditional static systems for clinically relevant applications. Biotechnology & Bioengineering 59:534–543.CrossRefGoogle Scholar
  32. Coutinho LH, Testa NG, Chang J, Morgenstern G, Harrison C, Dexter TM. 1990. The use of cultured bone marrow cells in autologous transplantation. Progress in Clinical and Biological Research 333:415–432.PubMedGoogle Scholar
  33. Craig W, Kay R, Cutler RL, Lansdorp PM. 1993. Expression of Thy-1 on human hematopoietic progenitor cells. Journal of Experimental Medicine 177:1331–1342.CrossRefPubMedGoogle Scholar
  34. Dao MA, Hashimo K, Kato I, Nolta JA. 1998. Adhesion to fibronectin maintains regenarative capacity during ex vivo culture and transduction of human hematopoietic stem and progenitor cells. Blood 92:4612–4621.PubMedGoogle Scholar
  35. De La Selle V, Gluckman E, Bruley-Rosset M. 1996. Newborn blood can engraft adult mice without inducing graft versus host disease across non H-2 antigens. Blood 87:3977–3984.Google Scholar
  36. Dexter TM, Allen TD, Lajtha LG. 1977. Conditions controlling the proliferation of haematopoietic stem cells in vivo. Journal of Cellular Physiology 91:335–344.CrossRefPubMedGoogle Scholar
  37. Douay L. 2001. Experimental Culture Conditions Are Critical for Ex Vivo Expansion of Hematopoietic Cells. Journal of Hematotherapy & Stem Cell Research 10:341–346.CrossRefGoogle Scholar
  38. Dybedal I, Jacobsen SE. 1995. Transforming growth factor beta (TGF-beta), a potent inhibitor of erythropoiesis: neutralizing TGF-beta antibodies show erythropoietin as a potent stimulator of murine burst-forming unit erythroid colony formation in the absence of a burst-promoting activity. Blood 86:949–957.PubMedGoogle Scholar
  39. Eaves CJ, Cashman JD, Kay RJ, Dougherty GJ, Otsuka T, Gaboury LA, Hogge DE, Lansdorp PM, Eaves AC, Humphries RK. 1991. Mechanisms that regulate the cell cycle status of very primitive hematopoietic cells in long-term human marrow cultures. II. Analysis of positive and negative regulators produced by stromal cells within the adherent layer. Blood 78:110–117.PubMedGoogle Scholar
  40. Eaves CJ, Eaves AC. 1997. Stem cell kinetics. Baillieres Clinical Haematology 10:233–257.Google Scholar
  41. Emmons RV, Doren S, Zujewski J, Cottler-Fox M, Carter CS, Hines K, O’shaughnessy JA, Leitman SF, Greenblatt JJ, Cowan K and others. 1997. Retroviral gene transduction of adult peripheral blood or marrow-derived CD34+ cells for six hours without growth factors or on autologous stroma does not improve marking efficiency assessed in vivo. Blood 89:4040–4046.PubMedGoogle Scholar
  42. Endo T, Ishibashi, Y., Okana, H., Fukumaki, Y. 1994. Sgnificance of pH on differentiation of human erythroid cell lines. Leukemia Research 18:49–54.CrossRefPubMedGoogle Scholar
  43. Erslev A, et al. 1983. Structure and function of the marrow. al. WWe, editor: New York: McGraw-Hill. 75-83 p.Google Scholar
  44. Fischkoff S, Pollak, A., Gleich, GJ., Testa, JR., Misawa, S., Reber, TJ. 1984. Eosinophilic differentiation of the human promyelocytic leukemia cell line. Journal of Experimental medicine 160:179–196.CrossRefPubMedGoogle Scholar
  45. Fortunel N, Hatzfeld, A. and Hatzfeld, JA. 2000. Transforming growth factor-B: pleiotropic role in the regulation of hematopoiesis. Blood 96(6):2022–2036.PubMedGoogle Scholar
  46. Gartner S, Kaplan, HS. 1980. Long-term culture of human bone marrow cells. Proc. Natl. Acad. Sci. (USA) 77:4756–4759.PubMedGoogle Scholar
  47. Giarratana M-C, Kobari, L., Neildez Nguyen, TMA., Firat, H., Bouchet, S., Lopez, M., Gorin, N-C., Thierry, D., Douay, L. 1998. Cell culture bags allow a large extent of ex vivo expansion of LTC-IC and functional mature cells which can subsequently be frozen: interest for large-scale clinical appplication. Bone Marrow Transplantation 22(7):707–715.CrossRefPubMedGoogle Scholar
  48. Goodell M, Brose K, Paradis G, Conner A, Mulligan A. 1996. Isolation and functional properties of murine hematopoietic stem cells that are replicating in vivo. Journal of Experimental Medicine 183:1797–1806.CrossRefPubMedGoogle Scholar
  49. Guba S, Gottschalk, LR., Jing, YH., Mulligan, T., Emerson, SG. 1992. Bone marrow stromal fibroblasts secrete interleukin-6 and granulocyte-macrophage colony-stimulating factor in the absence of inflammatory stimulation: demonstration by serum-free bioassay, enzyme-linked immunosorbent assay, and reverse transcriptase polymerase chain reaction. Blood 80(5):1190–1198.PubMedGoogle Scholar
  50. Haylock DN, To LB, Dowse TL, Juttner CA, Simmons PJ. 1992. Ex vivo expansion and maturation of peripheral blood CD34+ cells into the myeloid lineage. Blood 80:1405–1412.PubMedGoogle Scholar
  51. Hevehan D, Papoutsakis, ET., Miller, WM. 2000. Physiologically significant effect of pH and oxygen tension on granulopoiesis. Experimental Hematology 28:267–275.CrossRefPubMedGoogle Scholar
  52. Highfill J, Haley, SD., Kompala, DS. 1996. Large-scale production of murine bone marrow cells in an airlift packed bed bioreactor. Biotechnology and Bioengineering 50:514–520.CrossRefPubMedGoogle Scholar
  53. Holyoake TL, Freshney MG, McNair L, Parker AN, McKay PJ, Steward WP, Fitzsimons E, Graham GJ, Pragnell IB. 1996. Ex vivo expansion with stem cell factor and interleukin-11 augments both short-term recovery posttransplant and the ability to serially transplant marrow. Blood 87:4589–4595.PubMedGoogle Scholar
  54. Horner M, Miller ME, Ottino JM, Papoutsakis ET. 1998. Transport in a grooved perfusion flat-bed bioreactor for cell therapy applications. Biotechnology Progress 14:689–698.CrossRefPubMedGoogle Scholar
  55. Ishikawa Y, Ito, T. 1988. Kinetics of hematopietic stem cells in a hypoxic culture. European Journal of Haematology 40:126–129.PubMedGoogle Scholar
  56. Ivanovic Z, Bartolozzi, B., Bernabei, PA., et al. 2000. Incubation of murine bone marrow cells in hypoxia ensures the maintenance of marrow-repopulating ability together with the expansion of commited progenitors. British Journal of Haematology 108:424–429.CrossRefPubMedGoogle Scholar
  57. Ivanovic Z, Belloc, F., Faucher, J., et al. 2002. Hypoxia maintains and interleukin-3 reduces the pre-colony-forming cell potential of dividing CD34+ murine bone marrow. Experimental Hematology 30:67–73.CrossRefPubMedGoogle Scholar
  58. Jeevarajan A. 2002. Continuous pH Monitoring in a Perfused Bioreactor System Using an Optical pH Sensor. Biotechnology AND Bioengineering 78(4):467–472.CrossRefGoogle Scholar
  59. Kanai M, Hirayama, F., Yamaguchi, M., Ohkawara, J., Sato, N., Fukazawa, K., Yamashita, K., Kuwabara, M., Ikeda, H. & Ikebuchi, K. 2000. Stromal cell-dependent ex vivo expansion of human cord blood and augmentation of transplantable stem cell activity. Bone Marrow Transplantation 26:837–844.CrossRefPubMedGoogle Scholar
  60. Karanu FN, Murdoch B, Gallacher L, Wu DM, Koremoto M, Sakano S, Bhatia M. 2000. The notch ligand jagged-1 represents a novel factor of human hematopoietic stem cells. Journal of Experimental Medicine 192:1365–1372.CrossRefPubMedGoogle Scholar
  61. Kim D, Fujiki, Y., Fukushima, T., Ema, H., Shibuya, A., Nakauchi, H. 1999. Comparison of Hematopoietic Activities of Human Bone Marrow and Umbilical Cord Blood CD34 Positive and Negative Cells. Stem Cells 17:286–294.PubMedGoogle Scholar
  62. Kobayashi M, Laver JH, D LS, Kato T, Miyazaki H, Ogawa M. 1996. Thrombopoietin supports proliferation of human primitive hematopoietic cells in synergy with steel factor and/or interleukin-3. Blood 88:429–436.PubMedGoogle Scholar
  63. Kobayashi M, Laver JH, Lyman SD, Kato T, Miyazaki H, Ogawa M. 1997. Thrombopoietin, steel factor and the ligand for flt3/flk2 interact to stimulate the proliferation of human hematopoietic progenitors in culture. International. Journal of Hematology 66:423–434.CrossRefPubMedGoogle Scholar
  64. Koller M, Bender, JG., Papoutsakis, ET. & Miller, WM. 1992a. Effects of synergistic cytokine combinations, low oxygen, and irradiated stroma on the expansion of human cord blood progenitors. Blood 80(2):403–411.PubMedGoogle Scholar
  65. Koller M, Emerson, SG. & Palsson, BO. 1993. Large-Scale Expansion of Human Stem and Progenitor Cells From Bone Marrow Mononuclear Cells in Continuous Perfusion Cultures. Blood 82(2):378–384.PubMedGoogle Scholar
  66. Koller M, et al. 1992b. Reduced Oxygen Tension Increases Hematopoiesis in Long-term Culture of Human Stem and Progenitor Cells from Cord Blood and Bone Marrow. Experimental Hematology 20:264–270.PubMedGoogle Scholar
  67. Koller M, Manchel, I., Maher, RJ., Goltry, KL., Armostrong, D., Smith, AK. 1998. Clinical-Scale Human Umbilical Cord Blood Cell Expansion in a Novel Automated Perfusion Culture System. Bone Marrow Transplantation 21:653–663.CrossRefPubMedGoogle Scholar
  68. Koller M, Manchel, I., Palsson, MA., Maher, RJ. 1996. Different Measures of Human Hematopoietic Cell Culture Performance Are Optimized under Vastly Different Conditions. Biotechnology and Bioengineering 50:505–513.CrossRefPubMedGoogle Scholar
  69. Koller MR, Bender JG, Miller ME, Papoutsakis ET. 1993. Expansion of primitive human hematopoietic progenitors in a perfusion bioreactor system with IL-3, IL-6, and stem cell factor. Bio/Technology 11:358–63.CrossRefPubMedGoogle Scholar
  70. Koller MR, Bradley TR, Palsson BO. 1995a. Growth factor consumption and production in perfusion cultures of human bone marrow correlate with specific cell production. Experimental Hematology 23:1275–1283.PubMedGoogle Scholar
  71. Koller MR, Manchel I, Newsom BS, Palsson MA, Palsson BO. 1995b. Bioreactor expansion of human bone marrow: comparison of unprocessed, density-separated and CD34-enriched cells. Journal of Hematotherapy 4:159–169.PubMedGoogle Scholar
  72. Koller MR, Oxender M, Jensen TC, Goltry KL, Smith AK. 1999. Direct contact between CD34+/Lin-cells and stroma induces a soluble activity that specifically increases primitive hematopoietic cell production. Experimental Hematology 27:734–741.CrossRefPubMedGoogle Scholar
  73. Krause DS, Fackler MJ, Civin CI, May WS. 1996. CD34: structure, biology, and clinical utility. Blood 87:1–13.PubMedGoogle Scholar
  74. Krystal G, Lanm V, Dragowska W, Takahashi C, Appel J. 1994. Transforming growth factor beta 1 is an inducer of erythroid differentiation. Journal of Experimental Medicine 180:851–860.CrossRefPubMedGoogle Scholar
  75. Lansdorp PD, W. 1992. Long-Term Erythropoiesis from Constant Numbers of CD34+ Cells in Serum-free Cultures initiated with Highly Purified Progenitor Cells from Human Bone Marrow. Journal of Experimental Medicine 175:1501–1509.CrossRefPubMedGoogle Scholar
  76. Larochelle A, Vormoor J, Hanenberg H, Wang JC, Bhatia M, Lapidot T, Moritz T, Murdoch B, Xiao XL, Kato I and others. 1996. Identification of primitive human hematopoietic cells capable of repopulating NOD/SCID mouse bone marrow: implications for gene therapy. Nature Medicine 2:1329–1337.CrossRefPubMedGoogle Scholar
  77. Lill MC, Lynch M, Fraser JK, Chung GY, Schiller G, Glaspy JA, Souza L, Baldwin GC, Gasson JC. 1994. Production of functional myeloid cells from CD34-selected hematopoietic progenitor cells using a clinically relevant ex vivo expansion system. Stem Cells 12:626–637.PubMedCrossRefGoogle Scholar
  78. Lyman SD, Jacobsen SE. 1998. c-kit ligand and Flt3 ligand: stem/progenitor cell factors with overlapping yet distinct activities. Blood 91:1101–1134.PubMedGoogle Scholar
  79. Lyman SD, Williams DE. 1995. Biology and potential clinical applications for flt3 ligand. Current Opinion in Hematology 2:177–181.PubMedCrossRefGoogle Scholar
  80. Madlambayan GJ, Rogers I, Casper RF, Zandstra PW. 2001. Controlling Culture Dynamics for the Expansion of Hematopoietic Stem Cells. Journal of Hematotherapy & Stem Cell Research 10:481–492.CrossRefGoogle Scholar
  81. Mantalaris A, Keng P, Bourne P, Chang AYC, Wu JHD. 1998. Engineering a human bone marrow model: A case study on ex vivo erythropoiesis. Biotechnology Progress 14:126–133.CrossRefPubMedGoogle Scholar
  82. Martiat P, Ferrant, A., Cogneau, M., Bol, A., Rodhain, J., Michaux, JL., Sokal, G. 1987. Assessment of bone marrow blood flow using positron emmission tomography: No relationship with bone cellularity. Brithish Journal of Haematology 66:307–310.Google Scholar
  83. Martinson J, Unverzagt, K., Schaeffer, A., Smith, SL., Loudovaris, M., Schneidkraut, MJ., Bender, JG. and Van Epps, DE. 1998. Neutrophil Precursor Generation: Effect of Culture Conditions. Journal of Hematotherapy 7:463–471.PubMedGoogle Scholar
  84. Matsunaga T, Hirayama F, Yonemura Y, Murray R, Ogawa M. 1998. Negative regulation by interleukin-3 (IL-3) of mouse early B-cell progenitors and stem cells in culture: transduction of the negative signals by betac and betaIL-3 proteins of IL-3 receptor and absence of negative regulation by granulocyte-macrophage colony-stimulating factor. Blood 92:901–907.PubMedGoogle Scholar
  85. Mayani H. 1996. Composition and function of the hematopoietic microenvironment in human myeloid leukemia. Leukemia 10:1041–1047.PubMedGoogle Scholar
  86. Mayani HaL, PM. 1998. Biology of Human Umbilical Cord Blood-Derived Hematopoietic Stem/Progenitor Cells. Srem Cells 16:153–165.Google Scholar
  87. McAdams T, Miller, WM. & Papoutsakis, ET. 1996. Hematopoietic cell culture therapies (part I): cell culture considerations. Trends in Biotechnology 14:341–349.CrossRefPubMedGoogle Scholar
  88. McAdams T, Sandstrom, CE., Miller, WM., Bender, JG. and Paputsakis, ET. 1995. Ex vivo expansion of primitive hematopoietic cells for cellular therapies: An overview. Cytotechnology 18:133–146.CrossRefGoogle Scholar
  89. McAdams TA, Miller WM, Papoutsakis ET. 1997. Variations in culture pH affect the cloning eficiency and differentiation of progenitor cells in ex vivo haematopoiesis. British Journal of Haematology 97:889–895.CrossRefPubMedGoogle Scholar
  90. McAdams TA, Miller WM, Papoutsakis ET. 1998. pH is a potent modulator of erythroid differentiation. British Journal of Haematology 103:317–325.CrossRefPubMedGoogle Scholar
  91. McCune JM, Namikawa R, Kaneshima H, Shultz LD, Lieberman M, Weissman IL. 1998. The SCID-hu mouse: murine model for the analysis of human hematolymphoid differentiation and function. Science 24:1632–1639.Google Scholar
  92. McDowell CL, Papoutsakis ET. 1998. Increased agitation intensity increases CD13 receptor surface content and mRNA levels, and alters the metabolism of HL60 cells cultures in stirred tank bioreactors. Biotechnology & Bioengineering 60:239–250.CrossRefGoogle Scholar
  93. McNiece I, Briddell R. 2001. Ex vivo expansion of hematopoietic progenitor cells and mature cells. Experimental Hematology 29:3–11.CrossRefPubMedGoogle Scholar
  94. Meyer C, Drexler HG. 1999. FLT3 ligand inhibits apoptosis and promotes survival of myeloid leukemia cell lines. Leukemia Lymphoma 32:577–581.PubMedGoogle Scholar
  95. Mobest D, Goan, S., Junghahn, I., Winkler, J., Fichtner, I., Becker, M., De Lima-Hahn, E., Mertelsmann, R. & Henschler, R. 1999. Differential Kinetics of Primitive Hematopoietic Cells Assayed In Vitro and In Vivo During Serum-Free Suspension Culture of CD34+ Blood Progenitor Cells. Stem Cells 17:152–161.PubMedGoogle Scholar
  96. Mobest D, Mertelsmann, R. & Henschler, R. 1998. Serum-Free ex vivo Expansion of CD34+ Hematopoietic Progenitor Cells. Biotechnology and Bioengineering 60(3):341–347.CrossRefPubMedGoogle Scholar
  97. Mostafa S, Miller, WM & Papoutsakis, ET. 2000. Oxygen tension influences the differentiation, maturation and apoptosis of human megakaryocytes. British Journal of Haematology 111:879–889.CrossRefPubMedGoogle Scholar
  98. Naldini A, Carraro, F., Silvestri, S., et al. 1997. Hypoxia Affects Cytokine Production and Proliferation Responses by Human Peripheral Mononuclear Cells. Journal of Cellular Physiology 173:335–342.CrossRefPubMedGoogle Scholar
  99. Naughton BA, Jacob L, Naughton GK. 1990. A three-dimensional culture system for the growth of hematopoietic cells. Progress in Clinical and Biological Research 333:435–445.PubMedGoogle Scholar
  100. Naughton BA, Naughton GK. 1989. Hematopoiesis on nylon mesh templates-comparative long-term bone-marrow culture and the influence of stromal support cells. Annals of the New York Academy of Sciences 554:125–140.PubMedGoogle Scholar
  101. Naughton BA, Tjota A, Sibanda B, Naughton GK. 1991. Hematopoiesis on suspended nylon screen-stromal cell microenvironments. Journal of Biomechanical Engineering-Transactions of the ASME 113:171–177.Google Scholar
  102. Nielsen LK. 1999. Bioreactors for Hematopoietic Cell Culture. Annual Reviews in Biomedical Engineering 1:129–152.CrossRefGoogle Scholar
  103. Noll T, Jelinek, N., Schmidt, S., Biselli, M. & Wandrey, C. 2002. Cultivation of Hematopoietic Stem and Progenitor cells: Biochemical Engineering Aspects. Advances in Biochemical Engineering 74:111–128.Google Scholar
  104. Ogawa M, Clark SC. 1988. Synergistic interaction between interleukin-6 and interleukin-3 in support of stem cell proliferation in culture. Blood Cells 14:329–335.PubMedGoogle Scholar
  105. Oh D, Koller, MR. & Palsson, BO. 1994. Frequent Harvesting from Perfused Bone Marrow Cultures Results in in Increased Overall Cell and Progenitor Expansion. Biotechnology and Bioengineering 44:609–616.CrossRefPubMedGoogle Scholar
  106. Ohmizono Y, Sakaba H, Kimura T, Tanimukai S, Matsumura T, Miyazaki H, Lyman SD, Sonoda Y. 1997. Thrombopoietin augments ex vivo expansion of human cord blood-derived hematopoietic progenitors in combination with stem cell factor and flt3 ligand. Leukemia 11:524–530.CrossRefPubMedGoogle Scholar
  107. Osawa M, Hanada K, Hamada H, Nakauchi H. 1996. Long-term lymphohematopoietic reconstitution by CD34-low/negative hematopoietic stem cells. Science 273:242–245.PubMedGoogle Scholar
  108. Palsson BO, Paek S-H, Schwartz RM, Palsson MA, Lee G-M, Silver S, Emerson SG. 1993. Expansion of human bone marrow progenitor cells in a high cell density continuous perfusion system. Bio/Technology 11:368–372.CrossRefPubMedGoogle Scholar
  109. Pennathur-Das RL, L. 1987. Augmentation of In Vitro Human Marrow Erythropoiesis Under Physiological Oxygen Tensions Is Mediated by Monocytes and T Lymphocytes. Blood 69(3):899–907.PubMedGoogle Scholar
  110. Petzer AL, Zandstra PW, Piret JM, Eaves CJ. 1996. Differential cytokine effects on primitive (CD34+CD38-) human hematopoietic cells: novel responses to flt3-ligand and thrombopoietin. Journal of Experimental Medicine 183:2551–2558.CrossRefPubMedGoogle Scholar
  111. Piacibello W, Sanavio F, Garetto L, Severino A, Dane A, Gammaitoni I, Agglietta M. 1998. Differential growth factor requirement of primitive cord blood hematopoietic stem cells for self-renewal and amplification versus proliferation and differentiation. Leukemia 12:718–727.CrossRefPubMedGoogle Scholar
  112. Pierson BA, Europa AF, Hu WS, Miller JS. 1996. Production of human natural killer cells for adoptive immunotherapy using a computer-controlled stirred-tank bioreactor. Journal of Hematotherapy 5:474–483.Google Scholar
  113. Poloni A, Giarratana MC, Kobari L, Firat H, Bouchet S, Gorin NC, Douay L. 1997. The ex vivo expansion capacity of normal human bone marrow cells is dependent on experimental conditions: role of cell concentration, serum and CD34+ cell selection in stroma-free cultures. Hematological Cellular Therapy 39:49–58.CrossRefGoogle Scholar
  114. Quesenberry P, Colvin GA. 2001. Hematopoietic stem cells, progenitor cells, and cytokines. In: Seligsohn U, editor. Hematology. New York: McGraw-Hill. p 153–174.Google Scholar
  115. Quesenberry P, Crittenden R, Lowry P, Kittler E, Rao S, Peters S, Ramshaw H, Stewart F. 1994. In vitro and in vivo studies of stromal niches. Blood Cells 20:97–106.PubMedGoogle Scholar
  116. Rich I. 1986. A Role for the Macrophage in Normal Hemopoiesis. II. Effect of Varying Physiology Oxygen Tensions on the Release of Hemopoietic Growth Factors from Bone-marrow-derived Macrophage in vitro. Experimental Hematology 14:746–751.PubMedGoogle Scholar
  117. Rich I, & Kubanek, B. 1982. The effect of reduced oxygen tension on colony formation of erythropoietic cells in vitro. British Journal of Haematology 52:579–588.PubMedGoogle Scholar
  118. Rosenzwajg M, Canque B, Gluckman JC. 1996. Human dendritic cell differentiation pathway from CD34+ hematopoietic precursor cells. Blood 87:535–544.PubMedGoogle Scholar
  119. Rosenzweig M, Pykett MJ, Marks DF, Johnson RP. 1997. Enhanced maintenance and retroviral transduction of primitive hematopoietic progenitor cells using a novel three-dimensional culture system. Gene Therapy 4:928–936.CrossRefPubMedGoogle Scholar
  120. Rusten LS, Smeland EB, Jacobsen FW, Lien E, Lesslauer W, Loetscher H, Dubois CM, Jacobsen SE. 1994. Tumor necrosis factor-alpha inhibits stem cell factor-induced proliferation of human bone marrow progenitor cells in vitro. Role of p55 and p75 tumor necrosis factor receptors. Journal of Clinical Investigation 94:165–172.PubMedCrossRefGoogle Scholar
  121. Sandstrom CE, Bender JG, Miller ME, Papoutsakis ET. 1996. Development of a novel perfusion chamber to retain nonadherent cells and its use for comparison of human “mobilized” peripheral blood mononuclear cell cultures with and without irradiated bone marrow stroma. Biotechnology & Bioengineering 50:493–504.CrossRefGoogle Scholar
  122. Sandstrom CE, Bender JG, Papoutsakis ET, Miller WM. 1995. Effects of CD34+ cell selection and perfusion on ex vivo expansion of peripheral blood mononuclear cells. Blood 86:958–970.PubMedGoogle Scholar
  123. Sardonini CA, Wu YJ. 1993. Expansion and differentiation of human hematopoietic cells from static cultures through small-scale bioreactors. Biotechnology Progress 9:131–137.CrossRefPubMedGoogle Scholar
  124. Schwartz R, Palsson B, Emerson S. 1991. Rapid medium perfusion rate significantly increases the productivity and longevity of human bone marrow cultures. Proc. Natl. Acad. Sci. (USA) 88:6760–6764.PubMedGoogle Scholar
  125. Shah AJ, Smogorzewska EM, Hannum C, Crooks GM. 1996. Flt3 ligand induces proliferation of quiescent human bone marrow CD34+CD38 cells and maintains progenitor cells in vitro. Blood 87:3563–3570.PubMedGoogle Scholar
  126. Shih C-C, DiGiusto D, Forman SJ. 2002. Ex Vivo Expansion of Transplantable Human Hematopoietic Stem Cells: Where Do We Stand in the Year 2000? Journal of Hematotherapy & Stem Cell Research 9:621–628.Google Scholar
  127. Spangrude GJ, Johnson GR. 1990. Resting and activated subsets of mouse multipotent hematopoietic stem cells. Proceedings of the National Academy of Sciences USA 87:7433–7477.Google Scholar
  128. Sutherland HJ, Lansdorp PM, Henkelman DH, Eaves AC, Eaves CJ. 1990. Functional characterization of individual human hematopoietic stem cells cultured at limiting dilution on supportive marrow stromal layers. Proceedings of the National Academy of Sciences USA 87:3584–3588.Google Scholar
  129. Tennant G. 2000. Control of pH in human long-term bone marrow cultures with low-glucose medium containing zwitterion buffer lengthens the period of haematopoiesis activity. Brithish Journal of Haematology 109:785–787.CrossRefGoogle Scholar
  130. Till JE, McCulloch EA. 1961. A direct measurement of the radiation sensitivity of normal mouse bone marrow cells. Radiation Research 14:213–222.PubMedGoogle Scholar
  131. Trentin JJ. 1970. Influence of hematopoietic organ stroma (hematopoietic inductive microenvironments) on stem cell differentiation. In: Gordon AS, editor. Regulation of hemopoiesis. New York: Appleton-Century-crofts. p 161–186.Google Scholar
  132. Tsukada J, Misago, M., Kikuchi, M., Sato, T., Ogawa, R., Ota, T., Oda, S., Morimoto, I., Chiba, S. & Eto, S. 1992. Interaction between recombinant human erythropoietin and serum factor(s) on murine megakaryocyte colony formation. Blood 80(1):37–45.PubMedGoogle Scholar
  133. Veiby OP, Jacobsen FW, Cui L, Lyman SD, Jacobsen SE. 1996. The flt3 ligand promotes the survival of primitive hemopoietic progenitor cells with myeloid as well as B lymphoid potential. Suppression of apoptosis and counteraction by TNF-alpha and TGF-beta. Journal of Immunology 157:2953–2960.Google Scholar
  134. Verfaillie CM. 2001. Ex Vivo Expansion of Hematopoietic Stem Cells. In: Zon LI, editor. Hematopoiesis: A Developmental Approach. New York: Oxford University Press. p 119–129.Google Scholar
  135. Verfaillie CM, Hurley R, Bhatia R, McCarthy JB. 1994. MIP-1a combined with IL-3 conserves primitive human LTBMC-IC for at least 8 weeks in ex vivo “stroma-non-contact” cultures. Journal of Experimental Medicine 179:643–649.CrossRefPubMedGoogle Scholar
  136. Wang T-Y, Brennan JK, Wu JHD. 1995. Multilineal hematopoiesis in a three-dimensional murine longterm bone marrow culture. Experimental Hematology 23:26–32.PubMedGoogle Scholar
  137. Weissman I. 1994. Developmental switches in the immune system. Cell 76:207–218.CrossRefPubMedGoogle Scholar
  138. Williams DE, Hangoc G, Cooper S, Boswell HS, Shadduck RK, Gillis S, Waheed A, Urdal D, Broxmeyer HE. 1987. The effects of purified recombinant murine interleukin-3 and/or purified natural murine CSF-1 in vivo on the proliferation of murine hifg-and low-proliferative potential colony-forming cells: demonstration of in vivo synergism. Blood 70:401–403.PubMedGoogle Scholar
  139. Won JH, Cho SD, Park SK, Lee GT, Baick SH, Suh WS, Hong DS, Park HS. 2000. Thrombopoietin in synergy with other cytokines for expansion of cord blood progenitor cells. Journal of Hematotherapy and Stem Cell Research 9:465–473.CrossRefPubMedGoogle Scholar
  140. Yagi M, Ritchie KA, Sitnicka E, Storey C, Roth GI, Bartelmez S. 1999. Sustained ex vivo expansion of hematopoietic cells mediated by thrombopoietin. Proceedings of the National Academy of Sciences USA 96:8126–8131.CrossRefGoogle Scholar
  141. Yamaguchi M, Hiramaya, F., Kanai, M., Sato, N., Fukazawa, K., Yamashita, K., Sawada, K., Koike, T., Kuwabara, M., Ikeda, H. & Ikebuchi, K. 2001. Serum-free coculture system for ex vivo expansion of human cord blood primitive progenitos and SCID mouse-reconstituting cells using human bone marrow primary stromal cells. Experimental Hematology 29:174–182.CrossRefPubMedGoogle Scholar
  142. Yang H, Miller, WM., Papoutsakis, ET. 2002. High pH promotes megakaryocytic maturation and apoptosis. Stem Cells 20:320–328.CrossRefPubMedGoogle Scholar
  143. Zandstra PW, Conneally E, Petzer AL, Piret JM, Eaves CJ. 1997. Cytokine manipulation of primitive human hematopoietic cell self-renewal. Proceedings of the National Academy of Sciences USA 94:4698–4703.CrossRefGoogle Scholar
  144. Zandstra PW, Eaves CJ, Piret JM. 1994. Expansion of Hematopoietic Progenitor Cell Populations in Stirred Suspension Bioreactors of Normal Human bone Marrow Cells. Bio/Technology 12:909–914.CrossRefPubMedGoogle Scholar

Copyright information

© Springer 2005

Authors and Affiliations

  • L. Safinia
    • 1
  • N. Panoskaltsis
    • 2
  • A. Mantalaris
    • 1
  1. 1.Department of Chemical Engineering & Chemical TechnologyImperial College LondonLondonUK
  2. 2.Department of HaematologyImperial College LondonLondonUK

Personalised recommendations