Thrombopoietin to replace megakaryocyte-derived growth factor: impact on stem and progenitor cells during ex vivo expansion of CD34+ cells mobilized in peripheral blood

Transfusion ◽  
2010 ◽  
Vol 51 (2) ◽  
pp. 313-318 ◽  
Author(s):  
Pascale Duchez ◽  
Jean Chevaleyre ◽  
Marija Vlaski ◽  
Bernard Dazey ◽  
Fontanet Bijou ◽  
...  
Keyword(s):  
Growth Factor ◽  
Progenitor Cells ◽  
Peripheral Blood ◽  
Ex Vivo ◽  
Ex Vivo Expansion ◽  
Cd34 Cells ◽  
Stem And Progenitor Cells

scholarly journals Directed Differentiation of Mobilized Hematopoietic Stem and Progenitor Cells into Functional NK Cells with Enhanced Antitumor Activity

Cells ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 811
Author(s):  
Pranav Oberoi ◽  
Kathrina Kamenjarin ◽  
Jose Francisco Villena Ossa ◽  
Barbara Uherek ◽  
Halvard Bönig ◽  
...  
Keyword(s):  
Nk Cells ◽  
Progenitor Cells ◽  
Peripheral Blood ◽  
Cell Expansion ◽  
Nk Cell ◽  
Ex Vivo ◽  
Feeder Cells ◽  
Hematopoietic Stem ◽  
Cd34 Cells ◽  
Stem And Progenitor Cells

Obtaining sufficient numbers of functional natural killer (NK) cells is crucial for the success of NK-cell-based adoptive immunotherapies. While expansion from peripheral blood (PB) is the current method of choice, ex vivo generation of NK cells from hematopoietic stem and progenitor cells (HSCs) may constitute an attractive alternative. Thereby, HSCs mobilized into peripheral blood (PB-CD34+) represent a valuable starting material, but the rather poor and donor-dependent differentiation of isolated PB-CD34+ cells into NK cells observed in earlier studies still represents a major hurdle. Here, we report a refined approach based on ex vivo culture of PB-CD34+ cells with optimized cytokine cocktails that reliably generates functionally mature NK cells, as assessed by analyzing NK-cell-associated surface markers and cytotoxicity. To further enhance NK cell expansion, we generated K562 feeder cells co-expressing 4-1BB ligand and membrane-anchored IL-15 and IL-21. Co-culture of PB-derived NK cells and NK cells that were ex-vivo-differentiated from HSCs with these feeder cells dramatically improved NK cell expansion, and fully compensated for donor-to-donor variability observed during only cytokine-based propagation. Our findings suggest mobilized PB-CD34+ cells expanded and differentiated according to this two-step protocol as a promising source for the generation of allogeneic NK cells for adoptive cancer immunotherapy.

scholarly journals Mesenchymal Stem Cell-Derived Microvesicles Support Ex Vivo Expansion of Cord Blood-Derived CD34+Cells

2016 ◽  
Vol 2016 ◽  
pp. 1-13 ◽  
Author(s):  
Hui Xie ◽  
Li Sun ◽  
Liming Zhang ◽  
Teng Liu ◽  
Li Chen ◽  
...  
Keyword(s):  
Cord Blood ◽  
Progenitor Cells ◽  
Mononuclear Cells ◽  
Ex Vivo ◽  
Ex Vivo Expansion ◽  
Hematopoietic Stem ◽  
Promising Therapeutic Approach ◽  
Cd34 Cells ◽  
Stem And Progenitor Cells

Mesenchymal stem cells (MSCs) are known to support the characteristic properties of hematopoietic stem and progenitor cells (HSPCs) in the bone marrow hematopoietic microenvironment. MSCs are used in coculture systems as a feeder layer for the ex vivo expansion of umbilical cord blood (CB) to increase the relatively low number of HSPCs in CB. Findings increasingly suggest that MSC-derived microvesicles (MSC-MVs) play an important role in the biological functions of their parent cells. We speculate that MSC-MVs may recapitulate the hematopoiesis-supporting effects of their parent cells. In the current study, we found MSC-MVs containing microRNAs that are involved in the regulation of hematopoiesis. We also demonstrated that MSC-MVs could improve the expansion of CB-derived mononuclear cells and CD34+cells and generate a greater number of primitive progenitor cells in vitro. Additionally, when MSC-MVs were added to the CB-MSC coculture system, they could improve the hematopoiesis-supporting effects of MSCs. These findings highlight the role of MSC-MVs in the ex vivo expansion of CB, which may offer a promising therapeutic approach in CB transplantation.

Ex Vivo Expansion of Mobilized Peripheral Blood Myeloid Progenitor Cells: Pretreated Patients vs Normal Donors.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4135-4135
Author(s):  
Cécile De Bruyn ◽  
Alain Delforge ◽  
Dominique Bron
Keyword(s):  
Progenitor Cells ◽  
Peripheral Blood ◽  
Ex Vivo ◽  
Ex Vivo Expansion ◽  
Burst Activity ◽  
Myeloid Lineage ◽  
Myeloid Progenitor ◽  
Cd34 Cells ◽  
Myeloid Progenitor Cells ◽  
Pretreated Patients

Abstract Background: Additional infusion of ex vivo differentiated myeloid progenitor may reduce or abrogate severe neutropenia following high-dose chemotherapy and peripheral blood (PB) transplantation. Normal HPC as well as HPC from patients in clinical remission can be used as source of cells. The aims of our study were to compare the ex vivo expansion of myeloid progenitor cells starting from pretreated patients (PP) and normal donors (ND) and to evaluate if the cells collected from poor mobilizers -for which the ex vivo expansion is particularly interesting- present similar ex vivo expansion capacities than cells from good mobilizers. Methods: In this study, 11 normal donors (ND) and 17 patients (PP) were evaluated (5 NHL, 2 HK, 3 MM, 3 lymphoma, 1AML, 1 CLL and 2 solid tumor). The CD34+ cells were plated for 7 days in serum-free medium with SCF, IL-3, G-CSF and FLT3- ligand. The expression of CD34, CD13, CD15 and CD16 Ags and the presence of primary (myeloperoxydase, MPO) and secondary granules (lactoferrin, LF) were used to evaluate maturation into myeloid lineage. Functional capacities of the ex vivo generated cells were also analysed using oxydative burst activity and clonogenic capacities (CFU-GM). Results: (1) the proliferation capacities of total leucocytes were higher in ND than in PP (28.8±5.6 vs 17.2±4.0 fold, p<0.04) with a comparable CD34+ cell expansion (8.7±3.0 vs 7.8±2.7 fold, NS); (2) in patients, 65.3±5.8% of the expanded cells were CD13+ and 23.2±4.1% remained CD34+ with 7.8±2.5 % of immature CD34+CD13− cells; these results were similar to that observed in ND (72.7±7.5% CD13+, 23.0±5.3% CD34+, 9.0±3.2% CD34+CD13−); (3) the number of myeloid progenitors produced per seeded CD34+ cell consisted of 19.0±3.7 CD13+, 11.8±3.7 CD15+, 10.2±3.2 CD16+ and 15.4±4.4 MPO+/LF+cells in PP and 25.3±5.4 CD13+, 12.5±3.8 CD15+, 7.6±2.1 CD16+ and 21.7±7.2 MPO+/LF+ cells in ND (NS); (4) these cells, essentially promyelocytes and metamyelocytes, were able to proliferate in clonogenic assay and presented oxydative burst activity; (5) we failed to observe a correlation between the cell expansions (total leucocytes, CD34+ cells and myeloid progenitor cells) and the diagnosis; (6) no correlation was observed between the mobilization of CD34+ cells into PB and the capacities of cells to be expanded in vitro. Conclusions: in our culture system, the generation of cells committed to the myeloid lineage from CD34+ cells is feasible from ND as well as from PP, whatever is the diagnosis. The ex vivo expanded cell population consisted of cells at all stages of differentiation including immature hematopoietic progenitors. The mobilisation rate doesn’t correlate with the proliferative capacities of cells, suggesting that ex vivo expansion can be performed even in subjects with poor mobilization.

scholarly journals Ex vivo expansion of human hematopoietic stem and progenitor cells

Blood ◽  
2011 ◽  
Vol 117 (23) ◽  
pp. 6083-6090 ◽  
Author(s):  
Ann Dahlberg ◽  
Colleen Delaney ◽  
Irwin D. Bernstein
Keyword(s):  
Stem Cell ◽  
Growth Factors ◽  
Notch Signaling ◽  
Progenitor Cells ◽  
Ex Vivo ◽  
Ex Vivo Expansion ◽  
Hematopoietic Growth Factors ◽  
Self Renewal ◽  
Hematopoietic Stem ◽  
Stem And Progenitor Cells

AbstractDespite progress in our understanding of the growth factors that support the progressive maturation of the various cell lineages of the hematopoietic system, less is known about factors that govern the self-renewal of hematopoietic stem and progenitor cells (HSPCs), and our ability to expand human HSPC numbers ex vivo remains limited. Interest in stem cell expansion has been heightened by the increasing importance of HSCs in the treatment of both malignant and nonmalignant diseases, as well as their use in gene therapy. To date, most attempts to ex vivo expand HSPCs have used hematopoietic growth factors but have not achieved clinically relevant effects. More recent approaches, including our studies in which activation of the Notch signaling pathway has enabled a clinically relevant ex vivo expansion of HSPCs, have led to renewed interest in this arena. Here we briefly review early attempts at ex vivo expansion by cytokine stimulation followed by an examination of our studies investigating the role of Notch signaling in HSPC self-renewal. We will also review other recently developed approaches for ex vivo expansion, primarily focused on the more extensively studied cord blood–derived stem cell. Finally, we discuss some of the challenges still facing this field.

scholarly journals Maintenance of transplantation potential in ex vivo expanded CD34(+)- selected human peripheral blood progenitor cells

Blood ◽  
1994 ◽  
Vol 84 (9) ◽  
pp. 2898-2903 ◽  
Author(s):  
R Henschler ◽  
W Brugger ◽  
T Luft ◽  
T Frey ◽  
R Mertelsmann ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Peripheral Blood ◽  
Large Scale ◽  
Ex Vivo ◽  
Human Peripheral Blood ◽  
Ex Vivo Expansion ◽  
High Dose ◽  
Peripheral Blood Progenitor Cells ◽  
Limiting Dilution

Abstract CD34(+)-selected hematopoietic progenitor cells are being increasingly used for autotransplantation, and recent evidence indicates that these cells can be expanded ex vivo. Of 15 patients with solid tumors undergoing a phase I/II clinical trial using CD34(+)-selected peripheral blood progenitor cells (PBPCs) after high-dose chemotherapy, we analyzed the frequency of long-term culture-initiating cells (LTCIC) as a measure of transplantation potential before and after ex vivo expansion of CD34+ cells. PBPCs were mobilized by combination chemotherapy and granulocyte colony-stimulating factor (G-CSF). The original unseparated leukapheresis preparations, the CD34(+)-enriched transplants, as well as nonabsorbed fractions eluting from the CD34 immunoaffinity columns (Ceprate; CellPro, Bothell, WA) were monitored for their capacity to repopulate irradiated allogeneic stroma in human long-term bone marrow cultures. We found preservation of more than three quarters of fully functional LTCIC in the CD34(+)-selected fractions. Quantitation of LTCIC by limiting dilution analysis showed a 53-fold enrichment of LTCIC from 1/9,075 in the unseparated cells to an incidence of 1/169 in the CD34+ fractions. Thus, in a single apheresis, it was possible to harvest a median of 1.65 x 10(4) LTCIC per kg body weight (range, 0.71 to 3.72). In addition, in six patients, large-scale ex vivo expansions were performed using a five-factor cytokine combination consisting of stem cell factor (SCF), interleukin-1 (IL-1), IL-3, IL-6, and erythropoietin (EPO), previously shown to expand committed progenitor cells. LTCIC were preserved, but not expanded during the culture period. Optimization of ex vivo expansion growth factor requirements using limiting dilution assays for LTCIC estimation indicated that the five-factor combination using SCF, IL-1, IL-3, IL-6, and EPO together with autologous plasma was the most reliable combination securing both high progenitor yield and, at the same time, optimal preservation of LTCIC. Our data suggest that ex vivo-expanded CD34+ PBPCs might be able to allow long-term reconstitution of hematopoiesis.

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