Medium conditioned with mesenchymal stromal cell–derived osteoblasts improves the expansion and engraftment properties of cord blood progenitors

Dumont, Nellie; Boyer, Lucie; Émond, Hélène; Çelebi‐Saltik, Betül; Pasha, Roya; Bazin, Renée; Roy, Denis‐Claude; Pineault, Nicolas

doi:10.1016/j.exphem.2014.04.009

Cited by 17 publications

(54 citation statements)

References 51 publications

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“…The growth promoting activity of OCM is mediated by soluble and non-soluble factors We previously reported that 0.2 lm filtration reduced the capacity of OCM to support expansion of TNC (Dumont et al 2014). However, the impact of filtration on the growth and differentiation modulatory activities of OCM was not addressed further.…”

Section: Resultsmentioning

confidence: 98%

“…Interestingly, we previously showed that CB CD34 ? cells expanded in OCM improved platelet engraftment compared to cells expanded in standard SFM (Dumont et al 2014). However, the mean of action and active components of OCM responsible for this and other properties remained unresolved.…”

Section: Discussionmentioning

confidence: 99%

“…Human BM MSCs were established from purchased fresh BM mononuclear cells (Lonza, Walkersville, MD, USA) as previously described (Dumont et al 2014). A total of six plasticadherent MSC lines were generated from independent samples and used between passages 2-8.…”

Section: Methodsmentioning

confidence: 99%

“…Importantly, OCM was shown to modulate the engraftment properties of expanded HSPC. Indeed, human platelet levels were significantly greater in mice transplanted with HSPC expanded in OCM compared to control SFM or MSC CM (Dumont et al 2014).…”

Section: Introductionmentioning

confidence: 97%

“…Non-filtered centrifuged supernatant is then frozen without cryoprotectant until use (Dumont et al 2014). Herein, we sought to better understand and characterise the growth promoting activity of OCM on CB CD34 ?…”

Section: Introductionmentioning

confidence: 99%

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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: Resultsmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

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

et al. 2016

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Insights Into the Hematopoietic Regulatory Activities of Osteoblast by Secretomics

et al. 2020

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Osteoblasts are a key component of the endosteal hematopoietic stem cell niche and are recognized with strong hematopoietic supporting activity. Similarly, mesenchymal stromal cells (MSC)‐derived osteoblast (M‐OST) conditioned media (OCM) enhance the growth of hematopoietic progenitors in culture and modulate their engraftment activity. This article aims to characterize the hematopoietic supporting activity of OCM by comparing the secretome of M‐OST to that of their precursor. Over 300 proteins are quantified by mass spectroscopy in media conditioned with MSC or M‐OST, with 47 being differentially expressed. Growth factors, extracellular matrix proteins, and proteins from the complement pathways are included. The functional contribution of selected proteins on the growth and differentiation of cord blood (CB) progenitors is tested. Secreted protein acidic and rich in cysteine and Galectin 3 (Gal3) have little impact on the growth of CB cells in serum‐free medium (SFM). In contrast, inhibition of the complement 3A receptor (C3a‐R) present on CB progenitors significantly reduces the growth of CD34+ cells in OCM cultures but not in SFM. These results provide new insights into changes in factors released by MSC undergoing osteoblast differentiation, and on paracrine factors that are partially responsible for the hematopoietic supporting activity of osteoblasts.

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Paracrine Factors Released by Osteoblasts Provide Strong Platelet Engraftment Properties

et al. 2018

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Ex vivo expansion of hematopoietic stem cell (HSCs) and progenitors may one day overcome the slow platelet engraftment kinetics associated with umbilical cord blood transplantation. Serumfree medium conditioned with osteoblasts (i.e., osteoblast-conditioned medium [OCM]) derived from mesenchymal stromal cells (MSC) was previously shown to increase cell growth and raise the levels of human platelets in mice transplanted with OCM-expanded progenitors. Herein, we characterized the cellular and molecular mechanisms responsible for these osteoblast-derived properties. Limiting dilution transplantation assays revealed that osteoblasts secrete soluble factors that synergize with exogenously added cytokines to promote the production of progenitors with short-term platelet engraftment activities, and to a lesser extent with long-term platelet engraftment activities. OCM also modulated the expression repertoire of cell-surface receptors implicated in the trafficking of HSC and progenitors to the bone marrow. Furthermore, OCM contains growth factors with prosurvival and proliferation activities that synergized with stem cell factor. Insulin-like growth factor (IGF)-2 was found to be present at higher levels in OCM than in control medium conditioned with MSC. Inhibition of the IGF-1 receptor, which conveys IGF-2 0 intracellular signaling, largely abolished the growth-promoting activity of OCM on immature CD34 + subsets and progenitors in OCM cultures. Finally, IGF-1R effects appear to be mediated in part by the coactivator β-catenin. In summary, these results provide new insights into the paracrine regulatory activities of osteoblasts on HSC, and how these can be used to modulate the engraftment properties of human HSC and progenitors expanded in culture. STEM CELLS 2019;37:345-356 SIGNIFICANCE STATEMENTOsteoblasts are one of many different cell type implicated in the regulation of hematopoiesis through various mechanism including the release of paracrine factors such as growth factors, chemokines and extracellular matrix proteins. It has been shown that these factors strongly modulate the growth of umbilical cord blood stem and progenitor cells through activation of insulin-like growth factor-1 receptor and subsequent activation of the transcriptional activation complex β-catenin/TCF. Osteoblasts also modulated the expression repertoire of cell-surface receptors implicated in the homing of progenitors to the bone marrow. Altogether, these properties significantly enhanced the platelet engrafting activity of progenitors produced in cultures.

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Medium conditioned with mesenchymal stromal cell–derived osteoblasts improves the expansion and engraftment properties of cord blood progenitors

Cited by 17 publications

References 51 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

Insights Into the Hematopoietic Regulatory Activities of Osteoblast by Secretomics

Paracrine Factors Released by Osteoblasts Provide Strong Platelet Engraftment Properties

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