Rapid Expansion of Human Hematopoietic Stem Cells by Automated Control of Inhibitory Feedback Signaling

Csaszar, Elizabeth; Kirouac, Daniel C.; Yu, Mei; Wang, Weijia; Qiao, Wenlian; Cooke, M.; Boitano, Anthony E.; Ito, Caryn Y.; Zandstra, Peter W.

doi:10.1016/j.stem.2012.01.003

Cited by 227 publications

(243 citation statements)

References 32 publications

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“…CD38 -CD45RA -CD90 -HSC-like cells were significantly improved with the shorter 1 day conditioning than the original longer 6 days. Such finding suggest that longer conditioning period may result in the accumulation of factors such as MIP-1b, IL-8 and NAP-2 present in OCM (Dumont et al 2014) that promote the differentiation over the maintenance of immature cells such as MPP and HSC (Csaszar et al 2012). Besides, longer conditioning is associated with metabolic wastes buildup (e.g.…”

Section: Discussionmentioning

confidence: 98%

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: Discussionmentioning

confidence: 98%

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

et al. 2016

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“…These data clearly demonstrate that the bone marrow-on-a-chip faithfully mimics the natural physiological response of living bone marrow to clinically relevant doses of γ-radiation and to a validated radiation countermeasure drug (G-CSF), whereas conventional stroma-supported cultures do not. Importantly, the use of microfluidics also enables analysis of responses under flow, which is important for both regulation of marrow physiology 5,6,10,38,39 and study of pharmacokinetic and pharmacodynamic (PK/PD) behaviors of drugs that are critical for evaluation of their clinical behavior. …”

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confidence: 99%

Bone marrow–on–a–chip replicates hematopoietic niche physiology in vitro

et al. 2014

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____________________________________________________________________________The bone marrow microenvironment contains a complex set of cellular, chemical, structural and physical cues necessary to maintain the viability and function of the hematopoietic system [1][2][3][4][5] . This hematopoietic niche regulates hematopoietic stem cells (HSCs), facilitating a delicate balance between self-renewal and differentiation into progenitor cells that produce all mature blood cell types 4,5 . Engineering an artificial bone marrow that reconstitutes natural marrow structure and function, and that can be maintained in culture, could be a powerful platform to study hematopoiesis and test new therapeutics. It has proven difficult, however, to recreate the complex bone marrow microenvironment needed to support the formation and maintenance of a complete, functional hematopoietic niche in vitro [6][7][8][9] . Although various in vitro culture systems have (Revised NMETH-A19608) been developed to maintain and expand hematopoietic stem and progenitor cells 6-11 , there is currently no method to recreate or study the intact bone marrow microenvironment in vitro. Therefore, studies on hematopoiesis commonly rely on animal models to ensure the presence of an intact bone marrow microenvironment that enables normal physiologic marrow responses [12][13][14][15] . Furthermore, although it has been reported that bone marrow can be engineered in vivo [16][17][18][19] , no method exists to culture engineered bone marrow in vitro. To bridge the functional gap between in vivo and in vitro systems, we developed a method to produce a "bone marrow-on-a-chip" culture system that contains artificial bone and a living marrow, which is first generated in mice, and then explanted whole and maintained in vitro within a microfluidic device. RESULTS In vivo engineering of bone marrowTissue engineering methods have been used to induce formation of new bone with a central marrow compartment in vivo [18][19][20][21] . To explore the possibility of engineering an artificial bone marrow that can be explanted whole, we microfabricated a polydimethylsiloxane (PDMS) device with a central cylindrical cavity (1 mm high x 4 mm diameter) with openings at both ends (Fig. 1a). We filled the hollow compartment with a type I collagen gel containing bone-inducing demineralized bone powder (DBP) and bone morphogenetic proteins (BMP2 and BMP4) [20][21][22] , and implanted it subcutaneously on the back of a mouse (Supplementary Fig. 1). Our goal was to engineer bone that would fill the cylindrical space within the implanted device so that it could be easily removed whole and inserted into a microfluidic system containing a similarly shaped (Revised NMETH-A19608) chamber for in vitro culture (Fig. 1 a,b). These initial studies resulted in the creation of new bone encasing a marrow compartment that formed within the PDMS device 4 to 8 weeks after subcutaneous implantation. Histological analysis revealed that the marrow was largely inhabited by adipocytes and exhibite...

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“…Both of decreasing the concentration of accumulating negative secreted factors and increasing culture volume to maintain a lower cell density benefits HSC expansion. Using this system, a 12-d culture yielded an 11-fold increase of functional repopulating human cord blood HSCs relative to uncultured controls [120]. The perfusion and fed-batch systems thus represent unique and complementary approaches for expansion of HSCs through regulating the feedback signaling on HSC output.…”

Section: Continuous Perfusion and Fed-batch Systemsmentioning

confidence: 99%

“…A continuous perfusion system [119] and, more recently, a fed-batch system [120] designed to reduce the accumulating negative cues during the culture of HSCs significantly enhanced the expansion of functional primitive HSCs. Both of decreasing the concentration of accumulating negative secreted factors and increasing culture volume to maintain a lower cell density benefits HSC expansion.…”

Section: Continuous Perfusion and Fed-batch Systemsmentioning

confidence: 99%

“…An effective compound, UM171, was identified through further modification. In the Fed-Batch culture system [120], UM171 supports expansion of hematopoietic progenitors and results in a 13-fold expansion of SRCs. UM171 cooperates with SR1, an inhibitor of the AhR pathway, to induce an increase of hematopoietic progenitors in vitro; UM171, but not SR1, supports expansion of LT-HSCs.…”

Section: Um171mentioning

confidence: 99%

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Ex vivo expansion of hematopoietic stem cells

Xie

Zhang

2015

Sci. China Life Sci.

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Ex vivo expansion of hematopoietic stem cells (HSCs) would benefit clinical applications in several aspects, to improve patient survival, utilize cord blood stem cells for adult applications, and selectively propagate stem cell populations after genetic manipulation. In this review we summarize and discuss recent advances in the culture systems of mouse and human HSCs, which include stroma/HSC co-culture, continuous perfusion and fed-batch cultures, and those supplemented with extrinsic ligands, membrane transportable transcription factors, complement components, protein modification enzymes, metabolites, or small molecule chemicals. Some of the expansion systems have been tested in clinical trials. The optimal condition for ex vivo expansion of the primitive and functional human HSCs is still under development. An improved understanding of the mechanisms for HSC cell fate determination and the HSC culture characteristics will guide development of new strategies to overcome difficulties. In the future, development of a combination treatment regimen with agents that enhance self-renewal, block differentiation, and improve homing will be critical. Methods to enhance yields and lower cost during collection and processing should be employed. The employment of an efficient system for ex vivo expansion of HSCs will facilitate the further development of novel strategies for cell and gene therapies including genome editing. ex vivo expansion, hematopoietic stem cells, niche, signal transduction, cord blood, transplantation, SCID-repopulating cell, genome editing, CRISPR/Cas9 Citation:Xie JJ, Zhang CC. Ex vivo expansion of hematopoietic stem cells.

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Rapid Expansion of Human Hematopoietic Stem Cells by Automated Control of Inhibitory Feedback Signaling

Cited by 227 publications

References 32 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

Bone marrow–on–a–chip replicates hematopoietic niche physiology in vitro

Ex vivo expansion of hematopoietic stem cells

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