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Ex Vivo Expansion of Human Umbilical Cord Blood CD34+ Cells in a Collagen Bead—Containing 3-Dimensional Culture System

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Abstract

Self-renewal of stem cells depends on several critical factors, including the hematopoietic microenvironment, interactions with supporting stromal cells, features of the extracellular matrix, hematopoietic growth factors, and cytokines. Our study investigated the role of artificial 3-dimensional microenvironments as a means of replicating a more physiologic milieu in expansion of cord blood CD34+ cells. In the 3-dimensional model, hematopoietic cells inside collagen beads are exposed to cytokines added to a culture medium. We found that amplification of CD34+ cells with a clonogenic assay, fluorescenceactivated cell sorter analysis, and bone marrow repopulation of NOD/SCID mice showed greater clonogenic ability of cells cultured by the 3-dimensional method compared with the 2-dimensional method. The present study demonstrated that 3-dimensional matrix support may be useful for extended periods in expansion and preservation of stem cells or progenitor cells in vitro.

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References

  1. Kogler G, Callejas J, Hakenberg P, et al. Hematopoietic transplant potential of unrelated cord blood: critical issues.J Hematother. 1996;5:105–116.

    Article  PubMed  CAS  Google Scholar 

  2. Mayani H, Landsorp PM. Biology of human umbilical cord Blood—derived hematopoietic stem/progenitor cells.Stem Cells. 1998;16:153–165.

    Article  PubMed  CAS  Google Scholar 

  3. Kobari L, Pflumio F, Giarratana M-C, et al. In vitro and in vivo evidence for the long-term multilineage (myeloid, B, NK, and T) reconstitution capacity of ex vivo expanded human CD34+ cord blood cells.Exp Hematol. 2000;28:1470–1480.

    Article  PubMed  CAS  Google Scholar 

  4. Noort WA, Willemze R, Falkenburg JHF. Comparison of repopulating ability of hematopoietic progenitor cells isolated from human umbilical cord blood or bone marrow cells in NOD/SCID mice.Bone Marrow Transplant. 1998;22(suppl 1):S58-S60.

    PubMed  Google Scholar 

  5. Querol S, Capmany G, Cancelas JA, et al. Expansion of cord blood progenitor cells.Bone Marrow Transplant. 1998;21:S77-S80.

    PubMed  Google Scholar 

  6. Wagner JE, Kerman NA, Steinbuch M, et al. Allogeneic sibling umbilical-cord-blood transplantation in children with malignant and non-malignant disease.Lancet. 1995;346:214–219.

    Article  PubMed  CAS  Google Scholar 

  7. Locatelli F, Maccario R, Comoli P, et al. Hematopoietic and immune recovery after transplantation of cord blood progenitor cells in children.Bone Marrow Transplant. 1996;18:1095–1101.

    PubMed  CAS  Google Scholar 

  8. van Hennik PB, de Koning AE, Ploemacher RE. Seeding efficiency of primitive human hematopoietic cells in nonobese diabetic/ severe combined immune deficiency mice: implications for stem cell frequency assessment.Blood. 1999;94:3055–3061.

    PubMed  Google Scholar 

  9. Broxmeyer HE, Hangoc G, Cooper S, et al. Growth characteristics and expansion of human umbilical cord blood and estimation of its potential for transplantation in adults.Proc Natl Acad Sci U S A. 1992;89:4109–4113.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  10. Piacibello W, Sanavio F, Garetto L, et al. Differential growth factor requirement of primitive cord blood hematopoietic stem cell for self-renewal and amplificationvs proliferation and differentiation.Leukemia. 1998;12:718–727.

    Article  PubMed  CAS  Google Scholar 

  11. Kogler G, Callejas J, Sorg RV, et al. The effect of different thawing methods, growth factor combinations and media on the ex vivo expansion of umbilical cord blood primitive and committed progenitors.Bone Marrow Transplant. 1998;21:233–241.

    Article  PubMed  CAS  Google Scholar 

  12. Petzer AL, Zandstra PW, Piret JM, et al. Differential cytokine effects on primitive (CD34+CD38-) human hematopoietic cells: novel responses to Flt3-ligand and thrombopoietin.J Exp Med. 1996;183:2551–2558.

    Article  PubMed  CAS  Google Scholar 

  13. Piacibello W, Sanavio F, Garetto L, et al. Extensive amplification and self-renewal of human primitive hematopoietic stem cell from cord blood.Blood. 1997;89:2644–2653.

    PubMed  CAS  Google Scholar 

  14. Denning-Kendall PA, Nicol A, Horsley H, et al. Is in vitro expansion of human cord blood cells clinically relevant?Bone Marrow Transplant. 1998;21:225–232.

    Article  PubMed  CAS  Google Scholar 

  15. Li K, Liu J, Fok T, et al. Human neonatal blood: stem cell content, kinetics of CD34+ cell decline and ex vivo expansion capacity.Br J Haematol. 1999;104:178–185.

    Article  PubMed  CAS  Google Scholar 

  16. Xu R, Reems JA. Umbilical cord blood progeny cells that retain a CD34+ phenotype after ex vivo expansion have less engraftment potential than unexpanded CD34+ cells.Transfusion. 2001;41:213–218.

    Article  PubMed  CAS  Google Scholar 

  17. Lam AC, Li K, Zhang XB, et al. Preclinical ex vivo expansion of cord blood hematopoietic stem and progenitor cells: duration of culture; the media, serum supplements, and growth factors used; and engraftment in NOD/SCID mice.Transfusion. 2001;41:1567–1576.

    Article  PubMed  CAS  Google Scholar 

  18. Kohler T, Plettig R, Schaffer B, et al. Defining optimum conditions for the ex vivo expansion of human umbilical cord blood cells: influences of progenitor enrichment, interference with feeder layers, early-acting cytokines and agitation of culture vessels.Stem Cells. 1999;17:19–24.

    Article  PubMed  CAS  Google Scholar 

  19. Tun T, Miyoshi H, Ema H, et al. New type of matrix for bone marrow cell cultures: in vitro culture and in vivo transplantation experiments.ASAIO J. 2000;46:522–526.

    Article  PubMed  CAS  Google Scholar 

  20. Wang T-Y, Brennan JK, Wu JHD. Multilineal hematopoiesis in a three-dimensional murine long-term bone-marrow culture.Exp Hematol. 1995;23:26–32.

    PubMed  CAS  Google Scholar 

  21. Li Y, Ma T, Kniss DA, Yang ST, Laske LC. Human cord cell hematopoiesis in three dimensional nonwoven fibrous matrices: in vitro simulation of the marrow microenvironment.J Hematother Stem Cell Res. 2001;10:355–368.

    Article  PubMed  CAS  Google Scholar 

  22. Min YH, Lee ST, Choi KM, et al. Ex vivo generation of functional dendritic cells from mobilized CD34+ hematopoietic stem cells.Yonsei Med J. 1998;39:328–338.

    Article  PubMed  CAS  Google Scholar 

  23. Kie J-H, Yang W-I, Lee M-K, et al. Decrease in apoptosis and increase in polyploidization of megakaryocytes by stem cell factor during ex vivo expansion of human cord blood CD34+ cells using thrombopoietin.Stem Cells. 2002;20:73–79.

    Article  PubMed  CAS  Google Scholar 

  24. Ueda T, Tsuji K, Yoshino H, et al. Expansion of human NOD/ SCID-repopulating cells by stem cell factor, Flk2/flt3 ligand, thrombopoietin, IL-6, and soluble IL-6 receptor.J Clin Invest. 2000;105:1013–1021.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  25. DiGiusto DL, Lee R, Moon J, et al. Hematopoietic potential of cryopreserved and ex vivo manipulated umbilical cord blood progenitor cells evaluated in vitro and in vivo.Blood. 1996;84:1261–1271.

    Google Scholar 

  26. Kusadasi N, van Soest PL, Mayen AE, et al. Successful short-term ex vivo expansion of NOD/SCID repopulating ability and CAFC week 6 from umbilical cord blood.Leukemia. 2000;14:1944–1953.

    Article  PubMed  CAS  Google Scholar 

  27. Rossmanith T, Schroder B, Bug G, et al. Interleukin 3 improves the ex vivo expansion of primitive cord blood progenitor cells and maintains the engraftment potential of scid repopulating cells.Stem Cells. 2001;19:313–320.

    Article  PubMed  CAS  Google Scholar 

  28. Varas F, Bernard A, Bueren JA. Restrictions in the stem cell function of murine bone marrow grafts after ex vivo expansion of short-term repopulating progenitors.Exp Hematol. 1998;26:100–109.

    PubMed  CAS  Google Scholar 

  29. Holyoake T, Alcorn M, Richmond L, et al. CD34 positive PBPC expanded ex vivo may not provide durable engraftment following myeloablative chemoradiotherapy regimens.Bone Marrow Transplant. 1997;19:1095–1101.

    Article  PubMed  CAS  Google Scholar 

  30. Bentley SA. Close range cell:cell interaction required for stem cell maintenance in continuous bone marrow cultures.Exp Hematol. 1981;9:308–312.

    PubMed  CAS  Google Scholar 

  31. Szilvassy SJ, Bass MJ, van Zant G, et al. Organ-specific homing defines engraftment kinetics of murine hematopoietic stem cells and is compromised by ex vivo expansion.Blood. 1999;93:1557–1566.

    PubMed  CAS  Google Scholar 

  32. Koenigsmann MP, Koenigsmann M, Notter M, et al. Adhesion molecules on peripheral blood-derived CD34+ cells: effects of cryopreservation and short-term ex vivo incubation with serum and cytokines.Bone Marrow Transplant. 1998;22:1077–1085.

    Article  PubMed  CAS  Google Scholar 

  33. Peled A, Petit I, Kollet O, et al. Dependence of human stem cell engraftment and repopulation of NOD/SCID mice on CXCR4.Science. 1999;283:845–848.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Brain Korea 21 Project for Medical Sciences, USA

    Han-Soo Kim

  2. Departments of Internal Medicine, USA

    Han-Soo Kim, Yoo Hong Min & Seung Tae Lee

  3. Department of Laboratory Medicine, Yonsei University College of Medicine, Sinchon-Dong 134, Seodaemun-Ku, CPO Box 8044, Seoul, Korea

    Jong Baeck Lim & Hyun Ok Kim

  4. Pediatrics, Yonsei University, College of Medicine, Seoul

    Chuhl Joo Lyu

  5. Department of Oral and Maxillofacial Surgery, Chungnam National University Hospital, Daejeon, Korea

    Eun Seok Kim

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  1. Han-Soo Kim
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  2. Jong Baeck Lim
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  3. Yoo Hong Min
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  6. Eun Seok Kim
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  7. Hyun Ok Kim
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Correspondence to Hyun Ok Kim.

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Kim, HS., Lim, J.B., Min, Y.H. et al. Ex Vivo Expansion of Human Umbilical Cord Blood CD34+ Cells in a Collagen Bead—Containing 3-Dimensional Culture System. Int J Hematol 78, 126–132 (2003). https://doi.org/10.1007/BF02983380

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  • DOI: https://doi.org/10.1007/BF02983380

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