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1.
Seshadri T Al-Farsi K Stakiw J Ma C Saragosa R Franke N Keating A Crump M Kuruvilla J 《Bone marrow transplantation》2008,42(11):733-737
Peripheral blood hematopoietic progenitor cells (PBHC) are the standard source of support for high-dose chemotherapy because of faster recovery of marrow function. Unfortunately, a proportion of patients are unable to mobilize adequate progenitors to proceed to autologous hematopoietic cell transplant (AHCT). Granulocyte-CSF-stimulated BM-derived hematopoietic progenitor cells (BMHC) may circumvent this problem. From 1999 to 2006, 52 patients (cases) with AML, Hodgkin (HL) or non-Hodgkin's lymphoma (NHL) in whom PBHC mobilization failed underwent a G-CSF-stimulated bone marrow harvest and proceeded to AHCT. Their outcome was compared with 422 patients (controls) with AML, HL and NHL undergoing AHCT using only PBHC. Twenty-three patients received BMHC alone and 29 patients received a combination of PBHC and BMHC. Median engraftment time for neutrophils (>0.5 x 10(9)/l) and platelets (>20 x 10(9)/l) were 14 and 27 days, but significantly longer when compared with controls (11 days, 11 days, P<0.0001). Patients receiving both PBHC and BMHC had faster engraftment, when compared with those receiving BMHC alone (P<0.001). In conclusion, performing an AHCT using G-CSF-stimulated BMHC in patients failing PBHC collection is feasible with faster engraftment seen in patients receiving both BMHC and PBHC over BMHC alone. 相似文献
2.
J G Bender S F Williams S Myers D Nottleman W J Lee K L Unverzagt D Walker L B To D E Van Epps 《Bone marrow transplantation》1992,10(3):281-285
Twenty patients were treated with chemotherapy to mobilize progenitors into the blood. Peripheral blood stem cells were quantitated in peripheral blood or leukapheresis products using colony assays and flow cytometric measurement of CD34+ cells. In four patients where complete sets of serial samples were obtained, the appearance of CD34+ cells preceded the increase in CFU-GM by 24-48 h. Peak levels of CD34+ cells ranged from 0.6-5% and coincided with the peak increase in CFU-GM. Mobilized CD34+ cells contained subsets expressing CD33, CD13, CD45RA, CD38, HLA-DR, CD61 and CD41. Subsets of CD34+ cells expressing CD33, CD13, or CD45RA represent committed myeloid progenitors. In contrast to bone marrow CD34+ cells, few mobilized CD34+ cells expressed CD71, CD7, CD19 or CD10. Prompt engraftment of granulocytes greater than 500 x 10(6)/l at a median of 13 days and platelets greater than 50 x 10(9)/l at a median of 15 days was observed in patients reconstituted with mobilized cells. These data indicate that CD34+ cells mobilized during recovery from chemotherapy are predominantly myeloid in phenotype and contain few actively proliferating cells or cells with lymphoid phenotypes. 相似文献
3.
Differential maturation of megakaryocyte progenitor cells from cord blood and mobilized peripheral blood 总被引:7,自引:0,他引:7
Schipper LF Brand A Reniers N Melief CJ Willemze R Fibbe WE 《Experimental hematology》2003,31(4):324-330
OBJECTIVE: In comparison with stem cell transplantation using bone marrow or cytokine-mobilized peripheral blood, cord blood transplantation is characterized by delayed engraftment, in particular platelet recovery. The differences in the kinetics of engraftment may be related to quantitative differences in the numbers of stem cells and megakaryocyte progenitor cells and/or to qualitative differences between megakaryocyte progenitor cells in these grafts. We compared the hematopoietic composition of these grafts and determined the distribution of mature and immature megakaryocyte progenitor cells in cord blood and mobilized peripheral blood and their in vitro kinetic behavior. METHODS: Megakaryocyte progenitor cell subpopulations from cord blood (CB) and mobilized peripheral blood (PBSC) were expanded in vitro in the presence of mpl-ligand. The developmental differences during expansion of megakaryocyte progenitors were analyzed by flow cytometry and progenitor assays. RESULTS: We found that the immature (CD34(+)/CD41(-)) subpopulation from CB contains more than 98% of all megakaryocyte progenitor cells, responsible for 99% of all megakaryocytic cells cultured during 2 weeks. The CB CD34(+)/CD41(+) subpopulation shows no contribution to megakaryocytic cell formation. In contrast, in PBSC the mature (CD34(+)/CD41(+)) subpopulation contains 7% of all megakaryocyte progenitor cells. Moreover, CD34(+) cells from CB and PBSC also showed distinct phenotypic differences during maturation in vitro. PBSC megakaryocyte progenitor cells transiently express both CD34 and CD41 during maturation in vitro, whereas CB progenitor cells transiently lack expression of both markers before differention into (CD34(-)/CD41(+)) megakaryocytic cells. CONCLUSION: The in vitro data indicate the presence of different developmental stages of megakaryocyte progenitor cells in CB as compared to PBSC. These differences in composition and maturation between CB and PBSC may be related to the different kinetics of engraftment following transplantation of these stem cell sources. 相似文献
4.
Plastic-adherent progenitor cells in human bone marrow 总被引:2,自引:0,他引:2
Human bone marrow contains plastic-adherent hemopoietic progenitor cells whose plating efficiency is increased by brief (2 h) exposure to methylprednisolone (MP). When subsequently covered with methylcellulose medium, they form colonies of monoblastoid cells. Colony size, but not number, and mature cell production are increased by erythropoietin (epo) and granulocyte-macrophage colony-stimulating factor (GM-CSF). However, colonies do not grow under serum-free conditions. The resistance of plastic-adherent progenitors to treatment with 5-fluorouracil (5FU), their growth pattern, and their capacity to produce granulocytic and erythroid colonies on replating, suggest that they may be similar to the primitive, 5FU-resistant, plastic-adherent progenitor cells (HPP-CFC) in murine marrow. 相似文献
5.
Large-scale isolation of CD133+ progenitor cells from G-CSF mobilized peripheral blood stem cells 总被引:4,自引:0,他引:4
Gordon PR Leimig T Babarin-Dorner A Houston J Holladay M Mueller I Geiger T Handgretinger R 《Bone marrow transplantation》2003,31(1):17-22
We have evaluated the feasibility of large-scale isolation of CD133+ progenitors from healthy mobilized adult donors for potential clinical use in autologous and allogeneic transplantation. A total of 11 healthy volunteer adult donors were mobilized with G-CSF. CD133+ stem cells were isolated from a single leukapheresis using the Clinimacs method. The median percentage of CD133 before positive selection was 0.75% (range 0.39-2.03%). After selection, the median purity and recovery was 94% (range 85.2-98.0%) and 69% (range 44-100%), respectively. The median log10 T-cell depletion obtained by CD133+ positive selection was 4.2 (range 3.8-4.7). The CD133+ progenitors were highly enriched in colony-forming units (CFU) and transplantation into NOD/SCID mice resulted in a high engraftment rate. Transplantation of sorted CD133+/CD34+ cells into NOD/SCID mice showed a higher engraftment compared to CD133-/CD34+ cells. Mobilized peripheral CD133+ stem cells can be purified in large scale for potential clinical use. The biological function of the cells is not impaired. The majority of the NOD/SCID repopulating cells are within the CD133+/CD34+ subpopulation. Therefore, clinical studies using purified CD133+ stem cells can be envisoned to further clarify the role of CD133+ stem cells in hematopoietic reconstitution after transplantation. 相似文献
6.
Biological properties of peripheral blood progenitor cells mobilized by cyclophosphamide and granulocyte colony-stimulating factor 总被引:1,自引:0,他引:1
M. A. Scott J. F. Apperley D. M. Bloxham H. K. Jestice S. John R. E. Marcus & M. Y. Gordon 《British journal of haematology》1997,97(2):474-480
Patients transplanted with mobilized blood progenitor cells (PBPC) recover their neutrophil counts more rapidly than patients transplanted with bone marrow even when they receive the same dose/kg of granulocyte-macrophage colony-forming cells (CFU-GM). Here we have sought a biological explanation for this phenomenon. Most CD34-positive PBPC are quiescent (<1% in S phase) when they are collected from the bloodstream of patients treated with cyclophosphamide and granulocyte colony-stimulating factor (G-CSF), but we have shown that they are able to resume proliferation rapidly in vitro by measuring the kinetics of CFU-GM production by primitive plastic-adherent (PΔ) cells. Also, PΔcells in PBPC harvests, unlike normal marrow PΔ cells, were insensitive to cell-cycle restraint imposed by contact with marrow-derived stromal cells. We found that PΔ cells in PBPC collections produce relatively more CFU-GM and relatively fewer BFU-E than PΔ cells in bone marrow, indicating that granulopoiesis might occur at the expense of erythropoiesis, but we were unable to find any differences in the kinetics of granulocytic maturation between PBPC and bone marrow. Our interpretation of these findings is that transplanted PBPC rapidly enter the cell cycle and contact with stromal cells in the marrow does not reduce the proportion of progenitors participating in neutrophil production. Consequently, neutrophil recovery after PBPC infusion is more rapid than neutrophil recovery after marrow infusion. Granulopoiesis at the expense of erythropoiesis may also contribute to this effect. 相似文献
7.
Noncycling state of peripheral blood progenitor cells mobilized by granulocyte colony-stimulating factor and other cytokines 总被引:2,自引:8,他引:2
Incubation with high doses of tritiated thymidine in vitro was used to determine the percent of progenitor cells in the S phase of the cell cycle. Peripheral blood (PB), bone marrow (BM), and spleen populations from mice injected with granulocyte colony-stimulating factor (G-CSF) at 5 micrograms/day for 5 days and BM cells from uninjected littermates were assayed. Although the percentage of progenitor cells in S phase in the marrow (47% +/- 5%) and spleen (52% +/- 9%) was increased significantly in G-CSF-treated mice, only a small proportion of PB progenitor cells (PBPC) were in S phase (7% +/- 4%). In normal human subjects injected with G-CSF at 5 or 10 micrograms/kg/d, the proportions of PB myeloid (-1 +/- 4%) and erythroid (0% +/- 8%) progenitor cells in S phase were very low compared with the proportion of myeloid progenitor cells in S phase in normal BM (34% +/- 10%). Similarly, the large majority of steady-state PBPC and PBPC mobilized by interleukin-3 in combination with either granulocyte-macrophage colony-stimulating factor or G-CSF were also found not to be in S phase. Experiments indicated that the low percentages of PBPC in S phase were not ascribable either to inhibitory elements in the blood or to reduced responsiveness to growth factors. 相似文献
8.
Schiedlmeier B Wermann K Kühlcke K Eckert HG Baum C Fruehauf S Zeller WJ 《Bone marrow transplantation》2000,25(Z2):S118-S124
Mobilized peripheral blood progenitor cells (PBPC) are an attractive target for the retrovirus-mediated transfer of cytostatic drug resistance genes. We analyzed NOD/SCID mouse repopulating CD34+ PBPC from cancer patients following retroviral Transwell transduction in various cytokine combinations with the FMEV-based (Friend-mink cell focus forming/murine embryonic stem cell virus) hybrid vector SF-MDR carrying the human multidrug resistance-1 (MDR1) gene. Five to 10 weeks following transplantation of 2.0 x 10(6) CD34+ PBPC into NOD/SCID mice we observed medium to high levels of human cell engraftment with up to 33%. The extent of vector-marked human cells was assessed by a quantitative real-time polymerase chain reaction (PCR). SF-MDR gene transfer into long-term in vivo repopulating human hematopoietic cells was optimal in the presence of either IL-3/IL-6/SCF/FL or FL/TPO/SCF resulting in three-fold (12.4% +/- 1.7%) or four-fold (16.5% +/- 6.8%) higher average proportions of gene-marked human cells in NOD/SCID mice as compared to IL-3 alone (P < 0.01). In conclusion, we could optimize the engraftment capacity and the retroviral gene transfer to CD34+ PBPC using cocktails of early acting cytokines in combination with the recombinant fibronectin fragment CH-296. Our data suggest that the NOD/SCID model provides a valid assay to estimate the gene transfer efficiency to repopulating human PBPC that may be achievable in clinical autologous transplantation settings. 相似文献
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Giovanni D'Arena Pellegrino Musto Lazzaro Di Mauro Nicola Cascavilla Nicola Dello Iacono Potito Rosario Scalzulli Rosella Matera Mario Carotenuto 《American journal of hematology》1998,58(4):255-262
In order to investigate what is the best single parameter to predict the leukapheretic yield of circulating CD34+ progenitor cells, we retrospectively analyzed data from 68 patients with hematological malignancies who underwent mobilizing therapy. Three main parameters were monitored: total white blood cell (WBC), CD34+ cells, and monocyte counts in peripheral blood (PB) at the same day and at the preceding day of the apheretic procedure. Linear regression analysis revealed a strong correlation between CD34+ cell value in PB just before harvest and the number of CD34+ cells collected (P < 0.0001), but not at the preceding day. Monocyte PB concentration and absolute WBC count did not correlate with CD34+ cells harvested, at the preceding day of leukapheresis as well as at the same day of the procedure. The number of CD34+ cells in mobilized PB at the same day of harvest evidenced a very good capacity of predicting the value of harvested CD34+ cell number after collection, while WBC and monocyte count displayed quite a wide dispersion of results. In particular, an amount greater than 50/μL of circulating CD34+ cells ensured the best collections. Finally, CD34+ and CFU-GM content evaluated for each apheresis showed a strong reciprocal correlation (r 0.78; P < 0.0001). We conclude that the absolute number of CD34+ cells at the day of leukapheresis is the only parameter for identifying the exact timing for apheresis and predicting the amount of peripheral blood progenitor cells (PBPCs) that will be collected. In this setting, WBC and monocyte counts, at the day of collection or at the preceding day, are not useful tools. Am. J. Hematol. 58:255–262, 1998. © 1998 Wiley-Liss, Inc. 相似文献
11.
Lucien Perey Rowayda Peters Sandro Pampallona Philippe Schneider Nicole Gross & Serge Leyvraz 《British journal of haematology》1998,103(3):618-629
The transplantation of mobilized progenitor cells after high-dose chemotherapy shortens haemopoietic engraftment. CD34 cell subsets were examined in 20 consecutive mobilized progenitor cell collections obtained from patients with solid tumours that had not been previously treated. The analysis of CD34 cells was based on the expression of intracellular antigens, surface antigens including CD38, and cell size using multi-dimensional flow cytometry. We also correlated the numbers of stem cell subsets reinfused to haemopoietic recovery. The majority of CD34+ cells expressed CD13 and CD33. A significant proportion was cytoplasmic myeloperoxidase (cMPO) positive. CD34+ MPO+ cells increased significantly in late collections. MPO expression was related to cell size. Cells expressing CD13 also increased in late collections in parallel to CFU-GM count. Small subpopulations of CD34+ CD38+ were committed to B cells, T cells and erythroid cell lineages. A small population expressing the megakaryocytic antigen had a small size and were predominantly CD38?. A minor subpopulation expressed stem cells antigens. These were significantly higher in late collections (CD34+ Thy-1+ and CD34+ CD33?). After mobilization, patients received three cycles of intensive chemotherapy followed by reinfusion of mobilized progenitors (5.45 × 106/kg CD34+ cells, range 3.4–11.88). The numbers of reinfused CD34 cells or the individual subsets did not influence recovery of leucocytes (9 d) or platelets (9 d). In conclusion, the numbers of stem cells and their subsets differed between collections and, in unpretreated patients receiving intensive chemotherapy, there was no delayed engraftment when sufficient numbers of stem cells were reinfused. The recovery period was short and not correlated to any stem cell subsets. 相似文献
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Paola Omed Corrado Tarella Antonio Palumbo Chiara Argentino Daniele Caracciolo Paolo Corradini Alida Dominietto Fulvia Giaretta Roberta Ravaglia Roberta Triolo Sabrina Triolo Alessandro Pileri Mario Boccadoro 《British journal of haematology》1997,99(3):685-691
The possibility of reducing tumour cell contamination by cytotoxic drug courses prior to peripheral blood progenitor cell (PBPC) collection was evaluated in two consecutives groups of multiple myeloma (MM) patient candidates for autograft. All patients were at disease onset and received two VAD (vincristine, doxorubicin and dexamethasone) courses as initial debulking. In the first group (44 patients), mobilization and harvest were performed ‘upfront’, after a single cyclophosphamide (CY) administration of 4 g/m2; in the second group (17 patients), PBPC were collected at the end of a high-dose sequential chemotherapy programme, including: CY 5 g/m2, etoposide (VP16) 2 g/m2, a chemotherapy-free interval with three courses of high-dose dexamethasone, a final mobilizing CY at 7 g/m2. G-CSF was given following each high-dose cytotoxic drug. Cytofluorimetric analysis was performed to quantify progenitors (CD34+ cells) and plasma cells, identified by the high CD38 expression and/or CD38 and CD138 coexpression. Large amounts of PBPC were collected in either group (median harvested CD34+/kg: 15.8 × 106 and 13.4 × 106, respectively; P = 0.9). Circulating plasma cells were significantly higher in patients mobilized ‘upfront’ compared to those who received the high-dose sequence (median peak values of CD38bright/μl: 39 and 10, respectively; P = 0.02); a similar difference was observed in the amount of con-taminating plasma cells in the harvest products (median CD38bright/kg: 7.4 × 106 and 1.3 × 106, respectively; P = 0.02). The results demonstrate that an in vivo purging approach is feasible in myeloma patients through repeated high-dose chemotherapy courses; this may provide less-contaminated material suitable for further in vitro purging procedures. 相似文献
14.
Peripheral blood progenitor cell collections in multiple myeloma: predictors and management of inadequate collections 总被引:1,自引:0,他引:1
H. Miles Prince Kevin Imrie D. Robert Sutherland Armand Keating Jacinta Meharchand R. Michael Crump Caroline Girouard Kathy Trip A. Keith Stewart 《British journal of haematology》1996,93(1):142-145
Thirty-seven patients with previously treated multiple myeloma (MM) underwent peripheral blood progenitor cell (PBPC) collection following high-dose cyclophosphamide and GM-CSF or sequential IL-3 and GM-CSF. Patients with an inadequate collection were considered for a second or third collection. 25 patients underwent subsequent autotransplant. The only variable predictive of CFU-GM yield was the extent of prior melphalan therapy. All repeat collections were unsuccessful and patients infused with an autograft obtained from multiple sets of collections had a high incidence of delayed engraftment. We conclude that melphalan should be avoided or PBPC collection performed early in the disease course in patients who are potential transplant candidates. 相似文献
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16.
We have previously shown that administration of low-dose recombinant human stem cell factor (rhSCF) plus recombinant human granulocyte colony-stimulating factor (rhG-CSF) to baboons mobilizes greater numbers of progenitor cells in the blood than does administration of rhG-CSF alone. The purpose of the present study was to determine whether marrow repopulating cells are present in the blood of nonhuman primates administered low-dose rhSCF plus rhG-CSF, and if present, whether these cells engraft lethally irradiated recipients as rapidly as blood cells mobilized by treatment with rhG-CSF alone. One group of baboons was administered low-dose rhSCF (25 micrograms/kg/d) plus rhG- CSF (100 micrograms/kg/d) while a second group received rhG-CSF alone (100 micrograms/kg/d). Each animal underwent a single 2-hour leukapheresis occurring the day when the number of progenitor cells per volume of blood was maximal. For baboons administered low-dose rhSCF plus rhG-CSF, the leukapheresis products contained 1.8-fold more mononuclear cells and 14.0-fold more progenitor cells compared to the leukapheresis products from animals treated with rhG-CSF alone. All animals successfully engrafted after transplantation of cryopreserved autologous blood cells. In animals transplanted with low-dose rhSCF plus rhG-CSF mobilized blood cells, we observed a time to a platelet count of > 20,000 was 8 days +/- 0, to a white blood cell count (WBC) of > 1,000 was 11 +/- 1 days, and to an absolute neutrophil count (ANC) of > 500 was 12 +/- 1 days. These results compared with 42 +/- 12, 16 +/- 1, and 24 +/- 4 days to achieve platelets > 20,000, WBC > 1,000, and ANC > 500, respectively, for baboons transplanted with rhG-CSF mobilized blood cells. Animals transplanted with low-dose rhSCF plus rhG-CSF mobilized blood cells had blood counts equivalent to pretransplant values within 3 weeks after transplant. The results suggest that the combination of low-dose rhSCF plus rhG-CSF mobilizes greater numbers of progenitor cells that can be collected by leukapheresis than does rhG-CSF alone, that blood cells mobilized by low-dose rhSCF plus rhG-CSF contain marrow repopulating cells, and finally that using a single 2-hour leukapheresis to collect cells, the blood cells mobilized by low-dose rhSCF plus rhG-CSF engraft lethally irradiated recipients more rapidly than do blood cells mobilized by rhG- CSF alone. 相似文献
17.
The scarcity of donors for allogeneic bone marrow transplantation, the limited number of haematopoietic stem cell (HSC)/progenitors in cord blood samples and the sometimes insufficient number of mobilized peripheral blood cells collected from heavily treated cancer patients may benefit from ex vivo expansion of these cells for clinical transplantation. Depending on the clinical application, expansion of different haematopoietic cell subsets is required. HSC transplantation requires expansion of all cellular subsets including precursors, progenitors and HSCs for the short and long-term engraftment of patients. Quiescent HSCs may also be required for gene therapy by retrovirus. Finally, amplification of cells such as dendritic cells (DC) and different subsets of T and natural killer (NK) cells is required for immunotherapy. The different haematopoietic lineages are produced under different experimental conditions and the starting population is a critical parameter for the proposed clinical application. So it is essential to define the aims of haematopoietic cell expansion and to adapt the experimental conditions to obtain the required cell population. Mobilized peripheral blood cells are increasingly used as a source of haematopoietic cells. We review the biological characteristics of mobilized peripheral blood and the expansion of the different components according to the aims of their clinical use in the context of the progress currently achieved. 相似文献
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Mahmut N Katayama Y Takenaka K Teshima T Ohno Y Imajyo K Hara M Shinagawa K Ishimaru F Ikeda K Niiya K Harada M 《International journal of hematology》1999,69(1):36-42
We investigated the kinetics of posttransplant circulating progenitor cells (PTCPC) in the early phase after autologous (auto-) and allogeneic (allo-) peripheral blood stem cell transplantation (PBSCT). We analyzed the number of myeloid progenitor cells (CFU-GM) per 10 ml of peripheral blood (PB) on days 0 (just prior to transplantation), 1 (12-15 hours after completion of first transplantation), 2, 3, 5, 7, 10, 14, 17, 21 and 28 (after auto-PBSCT), and also additionally on day 35 after allo-PBSCT. A standard methylcellulose colony assay was used for analysing the number of CFU-GGM and BFU-E on all of the days. In addition, high proliferative potential-colony forming cells (HPP-CFC) of the harvested PBSC from donors and day 1 PB from recipients were assayed in 5 allo-PBSCT patients. Furthermore, a proportion of CD38- cells among CD34+ cells in the harvested PBSC and day 1 PB was evaluated by two-color flow cytometric analysis in 5 allo-PBSCT patients. The number of CFU-GM on day 1 ranged from 7 to 119 per 10 ml PB after auto-PBSCT, and from 15 to 61 per 10 ml PB after allo-PBSCT. After these transient increases, PTCPC diminished rapidly. Then, PTCPC emerged again on day 7 after auto-PBSCT and on day 10 or 14 after allo-PBSCT along with neutrophil recovery. A proportion of HPP-CFC among myeloid colonies from day 1 PB of recipients was significantly higher than that from the harvested PBSC from donors (65.6 +/- 12.7% vs. 17.4 +/- 13.0%, respectively, n = 5, P = 0.0013). In addition, two-color flow cytometric analysis revealed that the proportion of CD34+CD38- cells was significantly higher in day 1 PB of recipients than in the harvested PBSC from donors (57.5 +/- 17.6% vs. 11.7 +/- 4.9%, n = 5, P = 0.005). These observations suggest that both primitive and committed transplanted myeloid progenitor cells may circulate in the very early period following PBSCT. 相似文献
20.
Differences in cell cycle kinetics of candidate engrafting cells in human bone marrow and mobilized peripheral blood 总被引:4,自引:0,他引:4
Thornley I Nayar R Freedman MH Stephens D Crump M Messner HA Sutherland DR 《Experimental hematology》2001,29(4):525-533
Patients undergoing hematopoietic stem cell transplantation (HSCT) with mobilized peripheral blood (MPB) engraft quicker than those receiving bone marrow (BM). Our objective was to determine whether candidate engrafting cells--primitive hematopoietic progenitors (PHPs)--from MPB and BM exhibit different responses to cytokines that could explain this observation.We compared the cell cycle kinetics and ex vivo expansion of PHP-enriched cells obtained from MPB (n = 12) and BM (n = 10) by fluorescence-activated sorting of CD90+, AC133+ or CD38(dull) subsets of pre-selected CD34(+) cells. Cell cycle status, before and after 40 hours of serum-free culture with a cytokine cocktail, was assessed by multiparameter flow cytometry following incubation with Hoechst 33342 and pyronin Y.We found that 0.2% +/- 0.3% of MPB CD34(+)CD90(+) cells were in S/G(2)/M phases at hour 0, compared with 5% +/- 2.5% of those from BM (p = 0.0001), and 86.3% +/- 9.7% were in G(0), compared with 65.3% +/- 10% of those in BM (p = 0.0001). After 40 hours of culture, CD34(+)CD90(+) cells from MPB were more mitotically active than those from BM, with 29% +/- 4.9% in S/G(2)/M and 20% +/- 11.4% in G(0), compared to 19% +/- 6.5% (p = 0.001) and 39.2% +/- 22% (p = 0.027) of cells from BM. There was greater expansion of both total CD34(+) cells and the CD90(+) subset from MPB samples (p = 0.001 and 0.0001, respectively). Results from PHPs defined on the basis of AC133 expression correlated well with results obtained in CD90(+) subsets (r(2) = 0.81; p = 0.014).MPB PHPs appear to be primed for a greater acceleration in mitotic activity upon cytokine exposure. This qualitative difference may contribute to the earlier engraftment seen after HSCT using MPB grafts. 相似文献