Abstract
Ex vivo expansion of hematopoietic stem cells (HSCs) is very important for clinical applications of cord blood (CB). With the aim to find proper culture duration for ex vivo expansion, mononuclear cells (MNC) was applied as starting culture cells to expand HSCs and the repopulating potential of seven-day and fourteen-day cultured CD34+ cells were compared. The average expansion of total cells and CD34+ cells cultured for 7 days were higher than those cultured for 14 days. The results of phenotypic analysis of fresh and cultured cells showed that the percentage of CD3+ cells declined and the percentage of CD33+ cells increased during culture. The engraftment levels of fourteen-day cultured CD34+ cells were higher than those of fresh and seven-day cultured CD34+ cells. Fourteen-day cultured CD34+ cells also showed better multilineage reconstitution ability than fresh and seven-day cultured CD34+ cells. The results of the present study demonstrated that prolonged culture could preserve the hematopoietic reconstitution ability of ex vivo cultured CB cells and improve the engraftment level in NOD/SCID mice.
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Lane, T. A. (2005) Umbilical cord blood grafts for hematopoietic transplantation in adults: a cup half empty or half full? Transfusion 45: 1027–1034.
Robinson, S., T. Niu, M. de Lima, J. Ng, H. Yang, J. McMannis, S. Karandish, T. Sadeghi, P. Fu, M. del Angel, S. O’Connor, R. Champlin, and E. Shpall (2005) Ex vivo expansion of umbilical cord blood. Cytotherapy 7: 243–250.
Haylock, D. N., L. B. To, T. L. Dowse, C. A. Juttner, and P. J. Simmons (1992) Ex vivo expansion and maturation of peripheral blood CD34+ cells into the myeloid lineage. Blood 80: 1405–1412.
Douay, L. and G. Andreu (2007) Ex vivo production of human red blood cells from hematopoietic stem cells: what is the future in transfusion? Transfus. Med. Rev. 21: 91–100.
Matsunaga, T., I. Tanaka, M. Kobune, Y. Kawano, M. Tanaka, K. Kuribayashi, S. Iyama, T. Sato, Y. Sato, R. Takimoto, T. Takayama, J. Kato, T. Ninomiya, H. Hamada, and Y. Niitsu (2006) Ex vivo large-scale generation of human platelets from cord blood CD34+ cells. Stem Cells 24: 2877–2887.
Devine, S. M., H. M. Lazarus, and S. G. Emerson (2003) Clinical application of hematopoietic progenitor cell expansion: current status and future prospects. Bone Marrow Transplant. 31: 241–252.
Lam, A. C., K. Li, X. B. Zhang, C. K. Li, T. F. Fok, A. M. Chang, A. E. James, K. S. Tsang, and P. M. Yuen (2001) 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 41: 1567–1576.
Hexner, E. O., G. A. Danet-Desnoyers, Y. Zhang, D. M. Frank, J. L. Riley, B. L. Levine, D. L. Porter, C. H. June, and S. G. Emerson (2007) Umbilical cord blood xenografts in immunodeficient mice reveal that T cells enhance hematopoietic engraftment beyond overcoming immune barriers by stimulating stem cell differentiation. Biol. Blood Marrow Transplant. 13: 1135–1144.
Lapidot, T., Y. Fajerman, and O. Kollet (1997) Immunedeficient SCID and NOD/SCID mouse models as functional assays for studying normal and malignant human hematopoiesis. J. Mol. Med. 75: 664–673.
Rice, A. M., J. A. Wood, C. G. Milross, C. J. Collins, J. Case, M. R. Vowels, and R. E. Nordon (2001) Prolonged ex vivo culture of cord blood CD34+ cells facilitates myeloid and megakaryocytic engraftment in the non-obese diabetic severe combined immunodeficient mouse model. Br. J. Haematol. 114: 433–443.
Bhatia, M., D. Bonnet, U. Kapp, J. C. Wang, B. Murdoch, and J. E. Dick (1997) Quantitative analysis reveals expansion of human hematopoietic repopulating cells after short-term ex vivo culture. J. Exp. Med. 186: 619–624.
Yao, C. L., I. M. Chu, T. B. Hsieh, and S. M. Hwang (2004) A systematic strategy to optimize ex vivo expansion medium for human hematopoietic stem cells derived from umbilical cord blood mononuclear cells. Exp. Hematol. 32: 720–727.
Fietz, T., W. E. Berdel, H. Rieder, B. Reufi, H. Hopp, E. Thiel, and W. U. Knauf (1999) Culturing human umbilical cord blood: a comparison of mononuclear vs CD34+ selected cells. Bone Marrow Transplant. 23: 1109–1115.
Li, Q., H. Cai, Q. Liu, and W. S. Tan (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: 475–487.
Zandstra, P. W., C. J. Eaves, and J. M. Piret (1994) Expansions of hematopoietic progenitor cell populations in stirred suspension bioreactors of normal human bone marrow cells. Biotechnology 12: 909–914.
Araki, H., N. Mahmud, M. Milhem, R. Nunez, M. Xu, C. A. Beam, and R. Hoffman (2006) Expansion of human umbilical cord blood SCID-repopulating cells using chromatin-modifying agents. Exp. Hematol. 34: 140–149.
Ishikawa, F., A. G. Livingston, J. R. Wingard, S. Nishikawa, and M. Ogawa (2002) An assay for long-term engrafting human hematopoietic cells based on newborn NOD/SCID/β2- microglobulinnull mice. Exp. Hematol. 30: 488–494.
Xu, M. J., K. Tsuji, T. Ueda, Y. S. Mukouyama, T. Hara, F. C. Yang, Y. Ebihara, S. Matsuoka, A. Manabe, A. Kikuchi, M. Ito, A. Miyajima, and T. Nakahata (1998) Stimulation of mouse and human primitive hematopoiesis by murine embryonic aorta-gonadmesonephrosderived stromal cell lines. Blood 92: 2032–2040.
McNiece, I. K., G. Almeida-Porada, E. J. Shpall, and E. Zanjani (2002) Ex vivo expanded cord blood cells provide rapid engraftment in fetal sheep but lack long-term engrafting potential. Exp. Hematol. 30: 612–616.
Kobari, L., F. Pflumio, M. Giarratana, X. Li, M. Titeux, B. Izac, F. Leteurtre, L. Coulombel, and L. Douay (2000) 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. 28: 1470–1480.
Zhang, C., I. Todorov, Z. Zhang, Y. Liu, F. Kandeel, S. Forman, S. Strober, and D. Zeng (2006) Donor CD4+ T and B cells in transplants induce chronic graft-versus-host disease with autoimmune manifestations. Blood 107: 2993–3001.
Taussig, D. C., D. J. Pearce, C. Simpson, A. Z. Rohatiner, T. A. Lister, G. Kelly, J. L. Luongo, G. A. Danet-Desnoyers, and D. Bonnet (2005) Hematopoietic stem cells express multiple myeloid markers: implications for the origin and targeted therapy of acute myeloid leukemia. Blood 106: 4086–4092.
Kim, D. K., Y. Fujiki, T. Fukushima, H. Ema, A. Shibuya, and H. Nakauchi (1999) Comparison of hematopoietic activities of human bone marrow and umbilical cord blood CD34 positive and negative cells. Stem Cells 17: 286–294.
Ueda, T., K. Tsuji, H. Yoshino, Y. Ebihara, H. Yagasaki, H. Hisakawa, T. Mitsui, A. Manabe, R. Tanaka, K. Kobayashi, M. Ito, K. Yasukawa, and T. Nakahata (2000) 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. 105: 1013–1021.
Nitsche, A., I. Junghahn, S. Thulke, J. Aumann, A. Radonic, I. Fichtner, and W. Siegert (2003) Interleukin-3 promotes proliferation and differentiation of human hematopoietic stem cells but reduces their repopulation potential in NOD/SCID mice. Stem Cells 21: 236–244.
Levac, K., F. Karanu, and M. Bhatia (2005) Identification of growth factor conditions that reduce ex vivo cord blood progenitor expansion but do not alter human repopulating cell function in vivo. Haematologica 90: 166–172.
Madlambayan, G. J., I. Rogers, R. F. Casper, and P. W. Zandstra (2001) Controlling culture dynamics for the expansion of hematopoietic stem cells. J. Hematother. Stem Cell Res. 10: 481–492.
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Yang, S., Cai, H., Jin, H. et al. Hematopoietic reconstitution of CD34+ cells derived from short-term cultured cord blood mononuclear cells. Biotechnol Bioproc E 14, 429–435 (2009). https://doi.org/10.1007/s12257-008-0278-y
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DOI: https://doi.org/10.1007/s12257-008-0278-y