Journal of Hematology - Supplements - Haematologica

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24 the degree of lineage commitment. In freshly harvested CB we found that very few LTC-IC are in the S-phase of the cell cycle; however, in contrast to that which was observed in the bone marrow, also very few CFC were found in the S- phase of the cell cycle. A possible explanation is that a different regulation exists for bone marrow hematopoiesis and for CB hematopoiesis, but the factors which could determine this difference are presently unknown. Also, these authors found that cultured CD34 + cells reentering G0 have a shorter pre-replicating phase than freshly isolated G0 CD34 + cells. This observation could explain the high proportion (up to 90%, Figure 1) of LTC-IC and CFC entering the S-phase when incubated in the presence of cytokines for 36-48 hours. In summary, our data show that very few CBderived CFC and LTC-IC are in the S-phase of the cell cycle. Furthermore, results on cell cycle distribution and statin expression of CD34 + cells, while confirming the low numbers of progenitor cells in the S-phase of the cell cycle, clearly indicate that a significant number of CD34 + cells are in the G1 phase of the cell cycle. Our results also provide new information on the kinetics of recruitment into the S-phase of the cell cycle of CB-derived CFC and LTC-IC, and on culture conditions which can modify their cycling status without reducing their numbers. These findings could play an important role in the engineering of CB-derived progenitor cells for gene therapy and in the designing of clinically relevant protocols for their ex vivo expansion. References 1. Gluckman E. The therapeutic potential of fetal and neonatal hematopoietic stem cells. N Engl J Med 1996; 335:1839-40. 2. Gluckman E, Broxmeyer HE, Auerbach AD, et al. Hematopoietic reconstitution in a patient with Fanconi`s anemia by means of umbilical-cord blood from an HLA-identical sibling. N Engl J Med 1989; 321: 1174-8. 3. Kurtzberg J, Laughlin M, Graham ML, et al. Placental blood as a source of hematopoietic stem cells for transplantation into unrelated recipients. N Engl J Med 1996; 335:157-70. 4. Broxmeyer HE, Hangoc C, Cooper S, et al. Growth characteristics and expansion of human umbilical cord blood and estimation of its potential for tranplantation in adults. Proc Natl Acad Sci USA 1992; 89:4109-13. 5. Piacibello W, Sanavio F, Garetto L, et al. Extensive amplification and self-renewal of human primitive hematopoietic stem cells from cord blood. Blood 1997; 89:2644-53. 6. Zandstra PW, Conneally E, Piret PM, Eaves CJ. Ontogeny-associated changes in the cytokine responses of primitive human hematopoietic cells. Br J Haematol 1998; 101:770-8. 7. Kohn DB, Weinberg KI, Nolta JA, et al. Engraftment of gene-modified umbilical cord blood cells in neonates with adenosine deaminase deficiency. Nature Med 1995; 1:1017-23. 8. Ponchio L, Conneally E, Eaves C. Quantitation of the quiescent fraction of long term culture initiating cells in normal humanblood and marrow and the kinetics of their growth factor-stimulated entry into S-phase in vitro. Blood 1995; 86:3314-21. 9. Ponchio L, Eaves C, Hogge D, et al. Both cycling and noncycling primitive progenitors continue to be mobilized into the circulation during the leukapheresis of patients pretreated with chemotherapy and G-CSF. Br J Haematol1995; 99:394-402. 10. Danova M, Aglietta M. Cytokine receptors, growth factors and cell cycle in human bone marrow and peripheral blood hematopoietic progenitors. Haematologica 1997; 82:622-39. 11. Lemoli R, Tafuri A, Fortuna A, et al. Cycling status of CD34+ cells mobilized into peripheral blood of healthy donors by recombinant human granulocyte colony stimulating factor. Blood 1997; 89:1189-96. 12. Dresch C, El Kebir N, Metral J, Karsdorf, A. Cytosine arabinoside as a suicide agent for human colony forming cells. Exp Hematol 1983; 11:187-92. 13. Lucotti C, Malabarba L, Rosti V, et al. Cell cycle distribution of cord blood-derived heamatopoietic progenitor cells and their recruitment into the S-phase of the cell cycle. Br J of Haematol 2000; 108:621-8. 14. Preisler HD, Epstein J. A comparison of two methods for determining the sensitivity of human myeloid colony-forming units to cytosine arabinoside. Br J Haematol 1981; 47:519-27. 15. Carlo Stella C, Cazzola M, Ganser A, et al. Synergistic antiproliferative effect of recombinant interferon γ with recombinant interferon a on chronic myelogenous leukemia haematopoietic progenitor cells (CFU- GEMM, CFU-Mk, BFU-E, CFU-GM). Blood 1988; 72: 1293-9. 16. Lemoine FM, Humphries MK, Abraham SDM, Kristal G, Eaves CJ. Partial characterization of a novel stromal cell-derived pre-B-cell growth factor active on normal and immortalized pre-B cells. Exp Hematol 1988; 16: 718-25. 17. Sutherland HJ, Eaves CJ, Eaves AC, Dragowska W, Lansdorp PM. Characterization and partial purification of human marrow cells capable of initiating long term haematopoiesis in vitro. Blood 1989; 74:1563- 9. 18. Sutherland HJ, Eaves CJ. Long-term culture of human myeloid cells. In: Culture of Hematopoietic cells (ed. by Freshney RI, Pragnell IB and Freshney MG), New York, 1994, Wiley, 139-62. 19. Wang E. A 57.000-mol-wt protein uniquely present in non-proliferating cells and senescent human fibroblasts. J Cell Biol 1985; 100:545-51. 20. Pellicciari C, Mangiarotti R, Bottone MG, Danova M, Wang E. Identification of resting cells by dual parameter flow cytometry of statin expression and DNA content. Cytometry 1995; 21:329-37. 21. Rosti V, Bergamaschi G, Lucotti L, et al. Oligodeoxynucleotides antisense to c-abl specifically inhibit entry into S-phase of CD34+ hematopoietic cells end their differentiation to granulocyte-macrophage progenitors. Blood 1995; 86:3387-93. 22. Mangiarotti R, Bottone MG, Danova M, Pellicciari C. Bivariate flow cytometric analysis of DNA content versus immunopositivity for ribonucleotide reductase M1 subunit in the cell cycle. Cytometry 1998; 32:78-85. 23. Cashman JD, Eaves AC, Raines EW, et al. Mechanisms that regulate the cell cycle status of very primitive hematopoietic cells in long term human marrow cultures. I. Stimulatory role of a variety of mesenchi- haematologica vol. 85(supplement to n. 11):November 2000
25 mal cell activators and inhibitory role of TGFβ1. Blood 1990; 75:96-101. 24. Sitnicka E, Ruscetti FW, Priestle GV, Wolf NS, Bartelmez SH. Transforming growth factor b1 directly and reversibly inhibits the initial cell divisions of long term repopulating hematopoietic stem cells. Blood 1995; 88:82-8. 25. Sutherland HJ, Lansdorp PM, Henkelman JC, Eaves AC, Eaves JC. Functional characterization of individual human hematopoietic stem cells cultured in limiting dilution on supportive marrow stromal layers. Proc Natl Acad Sci USA 1990; 87:3584-8. 26. Movassagh M, Caillot L, Baillou C, Guigon M, Lemoine FM. Optimization of the cycling of clonogenic and primitive cord blood progenitor by various growth factors. Stem Cells 1997; 15:214-22. 27. Traycoff CM, Abboud MR, Laver J, Wade Clapp D, Srour EF. Rapid exit from G0/G1 phases of cell cycle in response to stem cell factor confers on umbilical cord blood CD34+ cells an enhanced ex vivo expansion potential. Exp Hematol 1994; 22:1264-72. 28. Hao QL, Shah AJ, ThiemannFT, Smogorzewska EM, Crooks GM. A functional comparison of CD34+ CD38– cells in cord blood and bone marrow. Blood 1995; 86:3745-53. 29. Roberts AW, Metcalf D. Noncycling status of peripheral blood progenitor cells mobilized by granulocyte colony-stimulating factor and other cytokines. Blood 1995; 86:1600-5. 30. Jordan CT, Yamasaki G, Minamoto D. High-resolution cell cycle analysis of defined phenotypic subsets within primitive human hemopoietic cell populations. Exp Hematol 1996; 24:1347-55. 31. Tanaka R, Katayama N, Ohishi K, et al. Accelerated cell-cycling of hematopoietic progenitor cells by growth factor. Blood 1995; 86:73-9. 32. Weeks SFA, Van Bockstaele DR, Plum J, et al. CD34++CD38– and CD34+CD38+ human hematopoietic progenitors from fetal liver, cord blood, and adult bone marrow respond differently to hematopoietic cytokines depending on the ontogenic source. Exp Hematol 1998; 26:1034-90. 33. Gothot A, Pyatt R, McMahel J, Rice S, Srour EF. Functional heterogeneity of human CD34+ cells isolated in subcompartments of the G0/G1 phase of the cell cycle. Blood 1997; 90:4384-92. haematologica vol. 85(supplement to n. 11):November 2000
Page 1 and 2: Haematologica established in 1920 e
Page 3 and 4: haematologica Journal of Hematology
Page 5 and 6: 3 Table 1. Indications for allogene
Page 7 and 8: 5 ease-free survival for childhood
Page 9 and 10: 7 Allogeneic bone marrow transplant
Page 11 and 12: haematologica 2000; 85(supplement t
Page 13 and 14: 11 achieved. However the relapse ra
Page 15 and 16: 13 ease. This observation is confir
Page 17 and 18: 15 plant failure. The role of DLI i
Page 19 and 20: 17 non-myeloablative regimen. Blood
Page 21 and 22: apheretic products obtained after m
Page 23 and 24: 21 Kinetics of activation of CFC in
Page 25: 23 according to the flow cytometric
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Page 31 and 32: 29 15 from activated cord versus ad
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Page 35 and 36: 33 Table 5. Timing and causes of tr
Page 37 and 38: 35 of children with storage disorde
Page 41 and 42: 39 parents wishes (the patient was
Page 45 and 46: 43 washing the cell suspension and
Page 47 and 48: 45 tive selection is usually around
Page 51 and 52: 49 chemotherapy blocks has been adm
Page 53 and 54: 51 Recently some clinical studies h
Page 55 and 56: 53 lymphoblastic leukaemia in Italy
Page 57 and 58: 55 Table 1. Summary of clinical dat
Page 59 and 60: 57 HLA-restricted cytomegalovirus (
Page 61 and 62: 59 that short-term artificial nutri
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Page 67 and 68: 65 References 1. Clarke DE, Raffin
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Page 71 and 72: 69 platin-thiotepa-etoposide is 34%
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75 Special Issues Conditioning The
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77 ticated studies with gene-marked
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79 treatment of poor risk systemic
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haematologica 2000; 85(supplement t
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83 Table 1. Rheumatologic disorders
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85 ly in terms of steroid-sparing a
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87 the same buffer was added to all
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haematologica 2000; 85(supplement t
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91 patients, 10 with a systemic for
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I index of authors Albertini A 41 A
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