Stromal factors support the expansion of the whole hemopoietic spectrum from bone marrow CD34+DR− cells and of some hemopoietic subsets from CD34+ and CD34+DR+ cells

Herman, P.; Ferrant, Augustin; Bruyère, Marc De; Straetmans, Nicole

doi:10.1038/sj.leu.2400924

Cited by 17 publications

(16 citation statements)

References 15 publications

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“…GF2, and we found that stromal cells significantly improve the expansion of CD34+ cells. These results are similar to those seen by others [12]. Except for one sample showing just a 5-fold expansion, which may be due to some technical errors in the processing of the sample, the remaining 9 samples showed significantly higher expansions ranging from 17.6- to 32-fold of CD34+ cells.…”

Section: Discussionsupporting

confidence: 80%

Ex vivo Expansion of Umbilical Cord Blood Stem Cells Using Different Combinations of Cytokines and Stromal Cells

Madkaikar¹,

Ghosh²,

Gupta³

et al. 2007

Acta Haematol

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Umbilical cord blood is a promising source of hematopoietic stem cells (HSC) for allogeneic transplantation. However, graft rejection and delayed engraftment remain major limitations, both of which are related to a limited number of stem cells in the cord blood graft. Ex vivo expansion of HSC has been suggested as one of the ways of overcoming the challenges caused by a limited hematopoietic cell number from cord blood stem cell transplantation. In this study, we quantified and characterized an ex vivo expansion capacity of cord blood-derived HSC in a liquid culture system under different conditions. These conditions included: the combinations and concentrations of hematopoietic growth factors [stem cell factor (SCF), granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin (IL)-3, IL-6 and erythropoietin (EPO)], placental conditioning medium (PCM), and stromal cell support. During culture, the mean nucleated cell count, the mean CD34+ cell count, fold expansion, viability, clonogenic assays and immunophenotypic characterization were performed on day 0, day 7, day 12 and day 14 on the expanded cellular product. The maximum expansion was achieved using GF2 (SCF + IL-3 + GM-CSF) with stromal cell support. The mean CD34+ cell expansion on days 7 and 12 was 16.25- and 21.4-fold (5.2–32), respectively, and the mean nucleated cell expansion was 15.1- and 21-fold (18.1–23.2). The mean nucleated cell viability on day 12 was 87.9% (85.6–92.5). After 12 days, granulocyte-macrophage colony-forming units CFU-GEMM showed a 20.4-fold increase. A 21.4-fold increase in the CD34+ cells and a 20-fold increase in the CFU-GEMM should provide enough cells from a single cord blood unit to reduce the period of cytopenia after single unit cord blood transplantation. Even if there was some doubt about the long-term repopulating capacity of the expanded cells part of the collected umbilical cord cells (25%) could be expanded till day 12 after transplanting the major part (75%) of the collection.

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Section: Discussionsupporting

confidence: 80%

Ex vivo Expansion of Umbilical Cord Blood Stem Cells Using Different Combinations of Cytokines and Stromal Cells

Madkaikar¹,

Ghosh²,

Gupta³

et al. 2007

Acta Haematol

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“…Long-term bone marrow culture (LTBMC) appears to embody many of the features of hematopoietic cell regulation in vivo and closely resembles the environment in hematopoietic tissues [25,26]. In vitro studies have shown that the adherent layer cells, either spontaneously or after activation, produce a number of positive soluble factors capable of maintenance, survival, proliferation, differentiation, and extensive self-renewal of hematopoietic cells [27][28][29]. The fact that hematopoiesis can be maintained for several weeks makes the LTBMC an ideal model for investigating the modulating effects of new compounds on hematopoietic tissue disorders.…”

Section: Contents Lists Available At Sciencedirectmentioning

confidence: 99%

Immunohematopoietic modulation by oral β-1,3-glucan in mice infected with Listeria monocytogenes

Torello¹,

Queiroz²,

Oliveira³

et al. 2010

International Immunopharmacology

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“…3 More than 50% of human marrow or cord blood LTC-IC can be maintained when cultured in contact with AFT024 feeders. 11,14 As non-contact culture conditions or culture in conditioned medium may be more easily adapted to the clinical setting, we here tested if human marrow derived LTC-IC can be maintained in AFT024 non-contact conditions. CD34 + /HLA-DR − cells were cultured in transwells above AFT024 and M2-10B4 feeders for 5 weeks and the absolute frequency of LTC-IC maintained after 5 weeks culture was assessed by LDA on M2-10B4 feeders.…”

Section: Resultsmentioning

confidence: 99%

“…[1][2][3][4][5] We and others have shown that primitive LTC-IC and primitive lymphoid progenitors can be maintained in stroma-based cultures even without contact with the stromal feeders. [6][7][8][9][10][11] Up to 50% of LTC-IC are maintained without exogenous cytokines in transwells above primary human bone marrow stroma or the murine fibroblast line M2-10B4, for 8 weeks. 3 Addition of IL-3 and MIP-1␣ increases this to 100%.…”

Section: Introductionmentioning

confidence: 99%

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Leukemia

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Soluble factors produced by human marrow stroma or the murine marrow derived M2-10B4 cell line support ex vivo maintenance for 5-8 weeks of 50% of human long-term culture initiating cells (LTC-IC). As the AFT024 cell line supports LTC-IC cultured in contact conditions better than M2-10B4 feeders, we evaluated LTC-IC support in non-contact conditions above AFT024 feeders. We show that only 15% of LTC-IC were maintained for 5 weeks in AFT024 non-contact cultures (n = 6, P Ͻ 0.05). As AFT024-conditioned media added to M2-10B4 non-contact cultures did not inhibit LTC-IC maintenance, AFT024 cells do not secrete factors that inhibit LTC-IC growth. We next characterized heparan sulfate glycosaminoglycans (HS-GAGs) and cytokines produced by AFT024 cells, which are both required for LTC-IC maintenance in M2-10B4 non-contact cultures. The size and extent of O-sulfation of HS-GAGs in AFT024 and M2-10B4 conditioned medium were similar, indicating that absence of hematopoietic specific HS-GAGs is not responsible for the lack of hematopoietic in AFT024 non-contact cultures. Levels of 13 different cytokines secreted in AFT024-and M2-10B4-conditioned medium were similar. However, addition of human SCF, G-CSF, GM-CSF, LIF, MIP-1␣ and IL-6 in concentrations found in human marrow stroma-conditioned medium to AFT024 non-contact cultures increased LTC-IC-maintenance to 72% at 5 weeks. These cytokines improved LTC-IC maintenance in part through interaction with the progenitors and in part, through interaction with the AFT024 feeder. Thus, although LTC-IC maintenance is poor in AFT024 non-contact cultures, addition of human cytokines enhances LTC-IC maintenance in part through indirect effects on the AFT024 feeder. Characterization of known or novel growth factors secreted by AFT024 cells before and after cytokine stimulation may lead to the identification of cytokines that support growth of human hematopoietic stem cells.

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Stromal factors support the expansion of the whole hemopoietic spectrum from bone marrow CD34+DR− cells and of some hemopoietic subsets from CD34+ and CD34+DR+ cells

Cited by 17 publications

References 15 publications

Ex vivo Expansion of Umbilical Cord Blood Stem Cells Using Different Combinations of Cytokines and Stromal Cells

Ex vivo Expansion of Umbilical Cord Blood Stem Cells Using Different Combinations of Cytokines and Stromal Cells

Immunohematopoietic modulation by oral β-1,3-glucan in mice infected with Listeria monocytogenes

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