A Stro-1+ human universal stromal feeder layer to expand/maintain human bone marrow hematopoietic stem/progenitor cells in a serum-free culture system

Gonçalves, Raquel; Silva, Cláudia Lobato da; Cabral, Joaquim M. S.; Zanjani, Esmail D.; Almeida-Porada, Graça

doi:10.1016/j.exphem.2006.05.024

Cited by 59 publications

(50 citation statements)

References 30 publications

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“…Both growth factor secretions and cell-cell contact have been reported to be needed for the growthmodulatory activates of MSC on HSPC (Flores-Guzman et al 2013;Peled et al 2004;Amsellem et al 2003). Moreover, coculture of HSPC in MSC contact-based cocultures has been shown to improve human chimerism after transplantation (Ferreira et al 2012;Goncalves et al 2006;Walenda et al 2011).…”

Section: Introductionmentioning

confidence: 99%

Characterization of the growth modulatory activities of osteoblast conditioned media on cord blood progenitor cells

et al. 2016

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Engraftment outcomes are strongly correlated with the numbers of hematopoietic stem and progenitor cells (HSPC) infused. Expansion of umbilical cord blood (CB) HSPC has gained much interest lately since infusion of expanded HSPC can accelerate engraftment and improve clinical outcomes. Many novel protocols based on different expansion strategies of HSPC and their downstream derivatives are under development. Herein, we describe the production and properties of serum-free medium (SFM) conditioned with mesenchymal stromal cells derivedosteoblasts (OCM) for the expansion of umbilical CB cells and progenitors. After optimization of the conditioning length, we show that OCM increased the production of human CB total nucleated cells and CD34 ? cells by 1.8-fold and 1.5-fold over standard SFM, respectively. Production of immature CD34 ? subpopulations enriched in hematopoietic stem cells was also improved with a shorter conditioning period. Moreover, we show that the growth modulatory activities of OCM on progenitor expansion are regulated by both soluble factors and non-soluble cellular elements. Finally, the growth and differentiation modulatory activities of OCM were fully retained after high dose-ionizing irradiation and highly stable when OCM is stored frozen. In summary, our results suggest that OCM efficiently mimics some of the natural regulatory activities of osteoblasts on HSPC and highlight the marked expansion potentials of SFM conditioned with osteoblasts.

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Section: Introductionmentioning

confidence: 99%

Characterization of the growth modulatory activities of osteoblast conditioned media on cord blood progenitor cells

et al. 2016

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“…We have previously shown that we can effectively expand CB‐HSPC and drive the differentiation of these cells toward both the myeloid and lymphoid lineages in a serum‐free culture system, using a feeder layer of adult human bone marrow (BM)‐derived stromal cells 4, 5, 6, 7. Using this system, we have optimized initial progenitor content and cytokine concentrations 7, 8, 9, and demonstrated that the expanded cells had the ability to engraft pre‐immune animals 5.…”

Section: Introductionmentioning

confidence: 99%

“…Using this system, we have optimized initial progenitor content and cytokine concentrations 7, 8, 9, and demonstrated that the expanded cells had the ability to engraft pre‐immune animals 5. Although the number of long‐term engrafting hematopoietic stem cells (HSC) amplified in this culture system, due to the overall increase in cell numbers, the absolute percentage growth of these most primitive stem cells decreased with time in culture.…”

Section: Introductionmentioning

confidence: 99%

Evaluating Interaction of Cord Blood Hematopoietic Stem/Progenitor Cells with Functionally Integrated Three-Dimensional Microenvironments

Mokhtari

Baptista

Vyas

et al. 2018

Stem Cells Translational Medicine

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Despite advances in ex vivo expansion of cord blood‐derived hematopoietic stem/progenitor cells (CB‐HSPC), challenges still remain regarding the ability to obtain, from a single unit, sufficient numbers of cells to treat an adolescent or adult patient. We and others have shown that CB‐HSPC can be expanded ex vivo in two‐dimensional (2D) cultures, but the absolute percentage of the more primitive stem cells decreases with time. During development, the fetal liver is the main site of HSPC expansion. Therefore, here we investigated, in vitro, the outcome of interactions of primitive HSPC with surrogate fetal liver environments. We compared bioengineered liver constructs made from a natural three‐dimensional‐liver‐extracellular‐matrix (3D‐ECM) seeded with hepatoblasts, fetal liver‐derived (LvSt), or bone marrow‐derived stromal cells, to their respective 2D culture counterparts. We showed that the inclusion of cellular components within the 3D‐ECM scaffolds was necessary for maintenance of HSPC viability in culture, and that irrespective of the microenvironment used, the 3D‐ECM structures led to the maintenance of a more primitive subpopulation of HSPC, as determined by flow cytometry and colony forming assays. In addition, we showed that the timing and extent of expansion depends upon the biological component used, with LvSt providing the optimal balance between preservation of primitive CB HSPC and cellular differentiation. Stem Cells Translational Medicine 2018;7:271–282

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“…We have previously reported [15,23] that CB-derived CD34 + cells could be expanded ex vivo in a MSC-based serum-free culture system containing SCF, FL, LIF and bFGF, differentiating primarily towards a myeloid phenotype, while maintaining a population of cells that expressed CD7, a marker of early lymphopoiesis. In this culture system, total CB CD34 + enriched cells expanded 124-358 fold, CD34+ cells increased by 35 fold, and CD34 + CD38 -cells by 48 fold by the end of culture.…”

Section: Differentiation Assaysmentioning

confidence: 99%

“…Cells were cultured in gelatin-coated T25 flasks with Mesenchymal Stem Cell Basal Medium (MSCBM ® ; Poietics™, Cambrex Bioscience, Baltimore, MD, USA) supplemented with MSCGM SingleQuot ® Kit. Stroma layers were cultured to confluence and then γ-irradiated with a 137 Cs source as previously described [15,23] Ex vivo expansion of CD34 + CD34 + cells were cultured in QBSF-60 serum-free medium with L-Glutamine (Quality Biological, Inc, Gaithersburg, MD, USA) supplemented with 100 ng/ml stem cell factor (SCF), 10 ng/ml leukemia inhibitor factor (LIF), 5 ng/ml basic fibroblast growth factor (bFGF) and 100 ng/ml Flt-3 ligand (FL) (all cytokines from PeproTech Inc., Rocky Hill, NJ, USA), on irradiated Stro-1 + stromal layers, at 37°C in a 5% (v/v) CO 2 incubator. Every 3 days, half of the medium was replaced with fresh medium and half the cultures were harvested for the following analyses: cell count, viability using trypan blue stain 0.4% solution (Gibco Laboratories, Grand Island, NY, USA), and phenotype by flow cytometry.…”

Section: Human Bone Marrow Stro-1 + Stroma Layer Cell Culturesmentioning

confidence: 99%

Generation of functional natural killer and dendritic cells in a human stromal-based serum-free culture system designed for cord blood expansion

Frias

Porada

Crapnell

et al. 2008

Experimental Hematology

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Objective-We have previously reported the ability of a mesenchymal stem cell (MSC)-based serum-free culture system to expand human cord blood (CB) hematopoietic stem cells (HSC) along the myeloid pathway and simultaneously generate a CD7 + CD34 -population. In this study, we investigated the ability of the CD7 + CD34 -population to differentiate into natural killer and dendritic cells.Materials and Methods-CB CD34 + cells were expanded over a MSC layer in serum-free medium supplemented with SCF, bFGF, LIF, and FL for 2 weeks. Cultured cells were harvested and CD7 + CD34 -Lin -cells sorted and plated for two additional weeks in either natural killer (NK)-or dendritic cell (DC)-inductive medium.Results-culture of CD34 + cells for the first 2 weeks in this system resulted in expansion of the stem cell pool and the myeloid component of the graft, and also produced a 58 fold-increase in the CD7 + CD34 -cell population. When sorted CD7 + CD34 -Lin -cells were induced towards a NK phenotype, further expansion was observed during this time in culture, and differentiation was confirmed by cytotoxic activity and by flow cytometry, with cells displaying CD16 and CD56 in the absence of CD3. The generation of DC cells in culture was also verified by observing both the characteristic dendritic morphology and the dendritic phenotypes HLA-DR bright CD123 bright CD11c -and HLA-DR bright CD11c + .Conclusion-These results demonstrate the ability of an ex-vivo culture system to drive the expansion of human CB HSCs while promoting the immune maturation of the graft and the generation of DC and NK cells that could then be utilized for adoptive cancer cellular immunotherapy.

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A Stro-1+ human universal stromal feeder layer to expand/maintain human bone marrow hematopoietic stem/progenitor cells in a serum-free culture system

Cited by 59 publications

References 30 publications

Characterization of the growth modulatory activities of osteoblast conditioned media on cord blood progenitor cells

Characterization of the growth modulatory activities of osteoblast conditioned media on cord blood progenitor cells

Evaluating Interaction of Cord Blood Hematopoietic Stem/Progenitor Cells with Functionally Integrated Three-Dimensional Microenvironments

Generation of functional natural killer and dendritic cells in a human stromal-based serum-free culture system designed for cord blood expansion

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