Characterization and transfusion of in vitro cultivated hematopoietic progenitor cells

Glück, Stefan; Chadderton, Tony; Porter, K.; Dietz, Günter; Maruyama, Midori

doi:10.1016/0955-3886(95)00035-v

Cited by 7 publications

(3 citation statements)

References 14 publications

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“…14 The number of nucleated cells infused was considerably less than the required estimate extrapolated from this series or from the computer-generated model. In other clinical trials of ex vivo cultured precursors, less committed progenitors were expanded and infused 22 and there was no appreciable impact on the duration of the neutropenic period.…”

Section: Discussionmentioning

confidence: 97%

“…12 Preclinical studies have also demonstrated that these cells, when maintained in culture, differentiate to fully functional neutrophils. 13 The feasibility of infusing expanded progenitors in the clinical setting has previously been reported, 14,15 but the clinical impact was less evident in these smaller series. We report here our experience with a larger group of patients with metastatic breast cancer who received ex vivo differentiated myeloid precursors as an adjunct to high-dose chemotherapy and peripheral blood progenitor cell rescue.…”

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confidence: 94%

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Clinical impact of ex vivo differentiated myeloid precursors after high-dose chemotherapy and peripheral blood progenitor cell rescue

Zimmerman

Lee

Bender

et al. 2000

Bone Marrow Transplant

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Summary:The infusion of ex vivo differentiated myeloid precursors may be able to shorten the period of obligatory neutropenia after high-dose chemotherapy and peripheral blood progenitor cell rescue by providing cells capable of differentiating to mature neutrophils within days of infusion. To test this hypothesis, 21 female patients with metastatic breast cancer underwent progenitor cell mobilization with cyclophosphamide, etoposide and G-CSF. CD34+ cells from one to two leukapheresis products were isolated and placed in suspension culture with a serum-free growth medium supplemented with PIXY321. The cultures were maintained for 12 days with subcultures initiated on day 7. The remaining leukapheresis products were cryopreserved in an unmanipulated state. Forty-eight hours after completing high-dose cyclophosphamide, thiotepa and carboplatin, the cryopreserved progenitors were infused, followed 1 to 24 h later by infusion of the differentiated myeloid precursors. In one patient, the cultured cells were labeled with Indium-111 with nuclear imaging performed up to 48 h post infusion. The differentiated myeloid precursors were suitable for infusion in 17 of the patients with a median 13-fold expansion of total nucleated cells. A range of 5.6 to 1066 × 10 7 nucleated cells were infused. Morphologically the cells were predominantly of myeloid lineage (63%) with a median 41% of the cells expressing CD15. No untoward effects were noted with the infusion of the cultured cells. The median days to neutrophil and platelet recovery were 8 and 10 days, respectively. There was a significant relationship (r = 0.67, P = 0.007) between the dose of differentiated myeloid precursors (CD15 + cells) and the depth and duration of neutropenia; a similar relationship, however, was also observed with the dose of cryopreserved CD34 + cells. After infusion of the radiolabeled myeloid precursors, a pattern of distribution similar to radio-labeled granulocytes was noted with uptake detected initially in the lungs and subsequently the reticulo-endothelial system. The impact of differentiated myeloid precursors on neutropenia as an adjunct to high-dose chemotherapy and peripheral blood pro-

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Section: Discussionmentioning

confidence: 97%

mentioning

confidence: 94%

Clinical impact of ex vivo differentiated myeloid precursors after high-dose chemotherapy and peripheral blood progenitor cell rescue

Zimmerman

Lee

Bender

et al. 2000

Bone Marrow Transplant

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“…Granulocytereduce or eliminate this neutropenia (9,10). Substantial macrophage-colony stimulating factor (GM-CSF) and numbers of these precursors can be obtained by culturgranulocyte-colony stimulating factor (G-CSF) either ing either bone marrow or mobilized peripheral blood alone (3)(4)(5) or in combination with peripheral blood pro-mononuclear cells (PBMC) (11) or selected CD34+ cells 'University of Chicago, Department of Medicine, Section of Hematology/Oncology, Chicago, IL 60637.…”

mentioning

confidence: 99%

Neutrophil Maturation of CD34+ Cells from Peripheral Blood and Bone Marrow in Serum-Free Culture Medium with PIXY321 and Granulocyte-Colony Stimulating Factor (G-CSF)

Smith

Bender

Berger

et al. 1997

Journal of Hematotherapy

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Bone marrow (BM) or peripheral blood (PB) CD34+ cells were cultured for 12 days in serum-free culture medium containing PIXY321 (IL-3/ GM-CSF fusion protein) with or without periodic supplements of granulocyte-colony stimulating factor (G-CSF). The cultures were evaluated at day 12 for total cell proliferation (fold increase from day 0), neutrophil differentiation by flow cytometry, using dual staining with CD15-FITC and CD11b-PE, and morphology using Wright-Giemsa and granule staining. In cultures containing PIXY321 where 6000 U/ml of G-CSF was added days 0 and 6, there was no significant difference (p > or = 0.05) in cell proliferation or the percent of CD15+/CD11b+ cells when compared with cultures with PIXY321 alone. ELISA analysis showed G-CSF levels had declined by 90% after 3 days of culture. Further studies were performed to assess the benefit of supplementing lower concentrations of G-CSF (600 U/ml) at more frequent intervals. A significant increase (p < or = 0.05) in cell proliferation and percent CD15+/CD11b+ was observed when G-CSF was added on days 0, 3, 6, and 9 (every 3 days) as compared with those cultures with PIXY321 alone. CD34+ cell proliferation without G-CSF was 19.6 +/- 4.8-fold, with G-CSF added on days 0 and 6 was 28.7 +/- 6.4-fold, and with G-CSF added on days 0, 3, 6, and 9 was 45.9 +/- 10.6-fold. Percent of CD15+/CD11b+ cells was 19.0 +/- 4.6%, 38.2 +/- 7.2%, and 58.5 +/- 6.5%, respectively, in these cultures. We observed more CD15+/CD11b+ cells, myelocytes/metamyelocytes, and secondary granule staining in cultures with G-CSF added on day, 0, 3, 6, and 9 as compared with cultures with G-CSF added on days 0 and 6 or no G-CSF added. We conclude that PIXY321 and G-CSF act synergistically on the in vitro proliferation and neutrophil differentiation of BM and PB CD34+ cells and that frequent supplements of G-CSF facilitate neutrophil differentiation.

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Ex vivo expansion of apheresis‐derived peripheral blood hematopoietic progenitors

Estrov¹,

Huh²,

Ginsberg³

et al. 2002

J of Clinical Apheresis

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Because the administration of hematopoietic growth factors and the use of stem cell support often fails to alleviate the neutropenic phase induced by cytotoxic drugs, several investigators have attempted to expand ex vivo hematopoietic progenitors for clinical use. These attempts have clearly shown that the cultured cells are functional and can be safely administered to patients, but that the in vivo performance is disappointing and the concept as a whole is not yet clinically useful. The major reasons for these unsuccessful attempts are thought to be cumbersome cell fractionation techniques, contamination, prolonged incubation, and the use of less than ideal cytokine combinations. In response, we have developed a simple procedure for ex vivo expansion of myeloid progenitor cells. In this assay, unfractionated mononuclear cells from apheresis donors are incubated in nonpyrogenic plastic bags for 7 days in the presence of culture medium either containing fetal calf serum or human plasma, granulocyte colony-stimulating factor, and stem cell factor. We have demonstrated that under these conditions the number of colony-forming units (CFU) granulocyte-macrophage (CFU-GM) and of CFU-granulocyte-macrophage-erythroid-megakaryocyte (CFU-GEMM) increased 7- and 9-fold, respectively, by day 7 and the number of burst-forming units-erythroid (BFU-E) increased 2.7-fold by day 5 of culture. Significant increases in the numbers of cells expressing CD34+, CD34+/CD38+, CD34+/CD33+, CD34+/CD15+, and CD34+/CD90+ and significant declines in the numbers of cells expressing CD34+/CD38- and CD19 surface antigens were also observed. The relative numbers of cells expressing T-cell markers and CD56 surface antigen did not change. By using different concentrations of various hematopoietic growth factor combinations, we can increase the number of mature and immature cells of different hematopoietic lineages.

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Characterization and transfusion of in vitro cultivated hematopoietic progenitor cells

Cited by 7 publications

References 14 publications

Clinical impact of ex vivo differentiated myeloid precursors after high-dose chemotherapy and peripheral blood progenitor cell rescue

Clinical impact of ex vivo differentiated myeloid precursors after high-dose chemotherapy and peripheral blood progenitor cell rescue

Neutrophil Maturation of CD34+ Cells from Peripheral Blood and Bone Marrow in Serum-Free Culture Medium with PIXY321 and Granulocyte-Colony Stimulating Factor (G-CSF)

Ex vivo expansion of apheresis‐derived peripheral blood hematopoietic progenitors

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