A novel protocol that allows short-term stem cell expansion of both committed and pluripotent hematopoietic progenitor cells suitable for clinical use

Astori, Giuseppe; Malangone, W.; Adami, Valentina; Risso, Angela; Dorotea, Laura; Falasca, E; Marini, L. D.; Spizzo, Riccardo; Bigi, Leonardo; Sala, Pierguido; Tonutti, Elio; Biffoni, Franco; Rinaldi, Cristina; Frate, G. Del; Pittino, Marco; Degrassi, A.

doi:10.1006/bcmd.2001.0439

Cited by 11 publications

(10 citation statements)

References 30 publications

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“…This finding is new and cannot be directly deduced from existing experimental data. Importantly, our model allows self-renewal of progenitor cells, as it is crucial after bone marrow transplantation (5,46,47).…”

Section: Discussionmentioning

confidence: 99%

Cell Division Patterns in Acute Myeloid Leukemia Stem-like Cells Determine Clinical Course: A Model to Predict Patient Survival

et al. 2015

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Acute myeloid leukemia (AML) is a heterogeneous disease in which a variety of distinct genetic alterations might occur. Recent attempts to identify the leukemia stem-like cells (LSC) have also indicated heterogeneity of these cells. On the basis of mathematical modeling and computer simulations, we have provided evidence that proliferation and self-renewal rates of the LSC population have greater impact on the course of disease than proliferation and self-renewal rates of leukemia blast populations, that is, leukemia progenitor cells. The modeling approach has enabled us to estimate the LSC properties of 31 individuals with relapsed AML and to link them to patient survival. On the basis of the estimated LSC properties, the patients can be divided into two prognostic groups that differ significantly with respect to overall survival after first relapse. The results suggest that high LSC self-renewal and proliferation rates are indicators of poor prognosis. Nevertheless, high LSC self-renewal rate may partially compensate for slow LSC proliferation and vice versa. Thus, model-based interpretation of clinical data allows estimation of prognostic factors that cannot be measured directly. This may have clinical implications for designing treatment strategies. Cancer Res; 75(6); 940-9. Ó2015 AACR.

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Section: Discussionmentioning

confidence: 99%

Cell Division Patterns in Acute Myeloid Leukemia Stem-like Cells Determine Clinical Course: A Model to Predict Patient Survival

et al. 2015

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“…Astori et al describe a novel protocol that enables short-term expansion of CD34 + cells for optimized production of early pluripotent hematopoietic SCs and committed cells from various lineages, and their strategy might represent a significant improvement for clinical application. 18 The application of ex-vivo SC expansion in cell-based therapy requires a standardized and well-controlled GMPcompliant methodology. The use of 6-well plates and bovine serum culture protocols allows adequate expansion levels with a high degree of reproducibility.…”

Section: Discussionmentioning

confidence: 99%

“…All these factors were chosen because they are early-acting cytokines that are proved to work on SCs according to the scientific literature. [18][19][20][21] Autologous cord blood plasma was taken from Ficoll gradients and peripheral blood plasma was obtained from 1 healthy donor. After 3 days 50% of the fresh medium was added to each well.…”

Section: Cell Expansionmentioning

confidence: 99%

Ex-Vivo Expanded Umbilical Cord Blood Stem Cells Retain Capacity for Myocardial Regeneration

Schlechta

Wiedemann

Kittinger

et al. 2010

Circ J

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Background: Umbilical cord blood (UCB) is a source of human hematopoietic precursor cells (HPCs), a stem cell (SC) type that has been used in several trials for myocardial repair. A certain minimal number of cells is required for measurable regeneration and a major challenge of SC-based regenerative therapy constitutes exvivo expansion of the primitive cell compartment. The aim of this study was to investigate the ex-vivo expansion potential of UCB-derived HPCs and the ability of these expanded cells to migrate to the site of damage and improve ventricular function in a rodent model of myocardial infarction (MI). Methods and Results:UCB-derived HPCs, defined by coexpression of CD133 and CD34, were expanded using various cytokine combinations. MI was induced by left anterior descending artery ligation in nude rats. Cells were injected intravenously 2 days after infarction. The combination of SC factor, thrombopoietin, flt3-ligand and interleukin-6 was found to be the most effective for inducing proliferation of HPCs. The migratory capacity of expanded HPCs was similar to that of non-expanded HPCs and improvement of ejection fraction was significant in both groups, with a relative increase of >60%.Conclusions: UCB-derived HPCs can be reproducibly expanded ex-vivo and retain their potential to improve cardiac function post-MI. (Circ J 2010; 74: 188 - 194)

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“…3,4 However, the currently defined culture protocols require long expansion periods, given the relative scarcity of blood DC precursors, and involve the use of extensive cytokine cocktails. [5][6][7][8] Therefore, a human cell line exhibiting the characteristics of CD34 ϩ -derived DC precursors would allow for the detailed study of DC differentiation without the associated problems of donor variability and DC precursor cell availability. It has been observed that cell lines derived from tumors of lymphoid or myeloid lineage may also share a potential for differentiation to DC-like APCs, thus providing a ready supply of DC precursors from which DCs can be easily and routinely generated.…”

mentioning

confidence: 99%

MUTZ-3, a human cell line model for the cytokine-induced differentiation of dendritic cells from CD34+precursors

Masterson

Sombroek

Gruijl

et al. 2002

Blood

143

145

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Many human myeloid leukemia-derived cell lines possess the ability to acquire a dendritic cell (DC) phenotype. However, cytokine responsiveness is generally poor, requiring direct manipulation of intracellular signaling mechanisms for differentiation. In contrast, the CD34 ؉ human acute myeloid leukemia cell line MUTZ-3 responds to granulocyte macrophagecolony-stimulating factor (GM-CSF), interleukin 4 (IL-4), and tumor necrosis factor alpha (TNF␣), cytokines known to be pivotal both in vivo and in vitro for DC generation from monocytes and CD34 ؉ stem cells. In all respects, MUTZ-3 cells behave as the immortalized equivalent of CD34 ؉ DC precursors. Upon stimulation with specific cytokine cocktails, they acquire a phenotype consistent with either interstitial-or Langerhans-like DCs and upon maturation (mDC), express CD83. MUTZ-3 DC display the full range of functional antigen processing and presentation pathways. These findings demonstrate the unique suitability of MUTZ-3 cells as an unlimited source of CD34 ؉ DC progenitors for the study of cytokineinduced DC differentiation. 3,4 However, the currently defined culture protocols require long expansion periods, given the relative scarcity of blood DC precursors, and involve the use of extensive cytokine cocktails. [5][6][7][8] Therefore, a human cell line exhibiting the characteristics of CD34 ϩ -derived DC precursors would allow for the detailed study of DC differentiation without the associated problems of donor variability and DC precursor cell availability. It has been observed that cell lines derived from tumors of lymphoid or myeloid lineage may also share a potential for differentiation to DC-like APCs, thus providing a ready supply of DC precursors from which DCs can be easily and routinely generated. However, many leukemia cell lines are often refractory to cytokine treatment, 9,10 requiring pharmacologic agents to induce a DC-like phenotype in myeloid cells, bypassing important checkpoints in the differentiation of DCs. 9,10 In contrast, it has been reported that the human cytokine-dependent myeloid cell line MUTZ-3 downregulates CD14 in response to interleukin 4 (IL-4) and low-level granulocyte macrophage-colony-stimulating factor (GM-CSF). 11,12 Here we demonstrate that this cell line is unique in its capacity to acquire a cytokine-induced interstitial and LC iDC phenotype, thus providing a rapid, logistically reproducible model for studies of the immunomodulatory capacity of DCs and such DC-related processes as antigen processing and presentation. Study design Generation of iDC-and mDC-like cells from leukemia cell linesThe cytokine-dependent human cell line MUTZ-3 (Deutsche Sammlung von Mikroorganismen und Zellkulturen [DSMZ], Braunschweig, Germany), and the cytokine-independent human cell lines, HL-60, KG-1, THP-1 U937, and K562 (American Type Culture Collection [ATCC], Manassas, VA) were cultured at 1 ϫ 10 5 /mL (total volume of 2.5 mL) in 12-well tissue culture plates (Costar, Cambridge, MA) in the presence of GM-CSF (100 ng/mL; Novartis/Scheri...

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A novel protocol that allows short-term stem cell expansion of both committed and pluripotent hematopoietic progenitor cells suitable for clinical use

Cited by 11 publications

References 30 publications

Cell Division Patterns in Acute Myeloid Leukemia Stem-like Cells Determine Clinical Course: A Model to Predict Patient Survival

Cell Division Patterns in Acute Myeloid Leukemia Stem-like Cells Determine Clinical Course: A Model to Predict Patient Survival

Ex-Vivo Expanded Umbilical Cord Blood Stem Cells Retain Capacity for Myocardial Regeneration

MUTZ-3, a human cell line model for the cytokine-induced differentiation of dendritic cells from CD34+precursors

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