Adhesion of synchronized human hematopoietic progenitor cells to fibronectin and vascular cell adhesion molecule-1 fluctuates reversibly during cell cycle transit in ex vivo culture

Huygen, Sandra; Giet, Olivier; Artisien, Vincent; Stefano, Ivano Di; Béguin, Yves; Gothot, André

doi:10.1182/blood.v100.8.2744

Cited by 21 publications

(22 citation statements)

References 41 publications

(37 reference statements)

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“…Control plates were coated with 1% bovine serum albumin (BSA; Sigma-Aldrich). 24 The coating solution was removed by aspiration and plates were incubated with RPMI 1640 (Cambrex Bio Science) containing 1% fraction V BSA at 37°C for 30 minutes to block nonspecific binding sites. After 2 washes in RPMI 1640 containing 25 mM HEPES (BioWhittaker Europe, Verviers, Belgium), 1 ϫ 10 5 to 2 ϫ 10 5 CD34 ϩ cells were plated in RPMI 1640 containing 0.1% BSA at 37°C in the presence of UTP, ATP, CXCL12, or control medium.…”

Section: Adhesion Assaymentioning

confidence: 99%

The extracellular nucleotide UTP is a potent inducer of hematopoietic stem cell migration

Rossi

Manfredini

Bertolini

et al. 2006

Blood

103

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Section: Adhesion Assaymentioning

confidence: 99%

The extracellular nucleotide UTP is a potent inducer of hematopoietic stem cell migration

Rossi

Manfredini

Bertolini

et al. 2006

Blood

103

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“…Current understanding of HSC biology implicates the roles of cell adhesion molecules N-cadherin and fibronectin in regulating stem cell quiescence and self-renewal [18]. The positive effects of surface-immobilized fibronectin in promoting expansion and maintenance of primitive characteristics of HSPCs have been demonstrated in several studies [10,11,[19][20][21][22][23]. In this study, nanofibers were not specifically modified with cell-adhesion ligands; and cells were cultured in serum-free medium.…”

mentioning

confidence: 99%

Functional nanofiber scaffolds with different spacers modulate adhesion and expansion of cryopreserved umbilical cord blood hematopoietic stem/progenitor cells

Chua

Chai

Lee

et al. 2007

Experimental Hematology

114

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Objective-Nanofiber scaffolds with amino groups conjugated to fiber surface through different spacers (ethylene, butylenes, and hexylene groups, respectively) were prepared and the effect of spacer length on adhesion and expansion of umbilical cord blood hematopoietic stem/progenitor cells (HSPCs) was investigated.Materials and Methods-Electrospun polymer nanofiber scaffolds were functionalized with poly (acrylic acid) grafting, followed by conjugation of amino groups with different spacers. HSPCs were expanded on aminated scaffolds for 10 days. Cell proliferation, surface marker expression, clonogenic potential, and nonobese diabetic (NOD)/severe combined immunodeficient (SCID) repopulation potential of the expanded cells were evaluated following expansion culture.Results-Aminated nanofiber scaffolds with ethylene and butylene spacers showed high-expansion efficiencies (773-and 805-fold expansion of total cells, 200-and 235-fold expansion of CD34 + CD45 + cells, respectively). HSPC proliferation on aminated scaffold with hexylene spacer was significantly lower (210-fold expansion of total cells and 86-fold expansion of CD34 + CD45 + cells), but maintained the highest CD34 + CD45 + cell fraction (41.1%). Colony-forming unit granulocyte-erythrocyte-monocyte-megakaryocyte and long-term culture-initiating cell maintenance was similar for HSPCs expanded on all three aminated nanofiber scaffolds; nevertheless, the NOD/SCID mice engraftment potential of HSPCs expanded on aminoethyl and aminobutyl conjugated nanofibers was significantly higher than that on aminohexyl conjugated nanofibers.Conclusion-This study demonstrated that aminated nanofibers are superior substrates for ex vivo HSPC expansion, which was correlated with the enhanced HSPC adhesion to these aminated nanofibers. The spacer, through which amino groups were conjugated to nanofiber surface, affected the expansion outcome. Our results highlighted the importance of scaffold topography and cellsubstrate interaction to regulating HSPC proliferation and self-renewal in cytokine-supplemented expansion.Offprint requests to: Hai-Quan Mao, Ph.D., Department of Materials Science and Engineering, Johns Hopkins University, 102 Maryland Hall, 3400 N. Charles Street, Baltimore, MD 21218; E-mail: hmao@jhu.edu. In conventional ex vivo expansion culture, HSPCs are generally regarded as suspension cells and numerous protocols implement HSPC suspension cultures in flasks or bags in the presence of various combinations of early acting cytokines [3][4][5][6][7]. The importance of topographical and substrate-immobilized biochemical cues has not been taken into consideration in these culture systems. Suspension culture conditions differ drastically from conditions provided by the natural bone marrow microenvironment, where a complex network of stromal cells and extracellular matrix (ECM) serves as a stem cell niche that regulates HSPC functions such as homing, self-renewal, proliferation, and fate choice [2,8]. A growing body of evidence has suggested the importance of...

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“…After 16 hours, aphidicolin (2 g/mL; Sigma, Poole, United Kingdom) was added, and incubation was continued for another 24 hours, as described previously. 26 At the end of this incubation (40 hours altogether), cells were washed and replated in X-VIVO 10/1% HSA with cytokines to allow S-phase progression. Cells were harvested at 3 and 6 hours thereafter to assess their position in the cell cycle and their homing ability.…”

Section: Cell Synchronization Using Aphidicolinmentioning

confidence: 99%

Impaired bone marrow homing of cytokine-activated CD34+ cells in the NOD/SCID model

Ahmed

Ings

Pizzey

et al. 2004

Blood

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The reduced engraftment potential of hematopoietic stem/progenitor cells (HSPCs) after exposure to cytokines may be related to the impaired homing ability of actively cycling cells. We tested this hypothesis by quantifying the short-term homing of human adult CD34 ؉ cells in nonobese diabetic/severe combined immunodeficient (NOD/SCID) animals. We show that the loss of engraftment ability of cytokine-activated CD34 ؉ cells is associated with a reduction in homing of colony-forming cells (CFCs) to bone marrow (BM) at 24 hours after transplantation (from median 2.8% [range, 1.9%-6.1%] to 0.3% [0.0%-0.7%]; n ‫؍‬ 3; P < .01), coincident with an increase in CFC accumulation in the lungs (P < .01). Impaired BM homing of cytokine-activated cells was not restored by using sorted cells in G 0 G 1 or by inducing cell cycle arrest at the G 1 /S border. Blocking Fas ligation in vivo did not increase the BM homing of cultured cells. Finally, we tested cytokine combinations or culture conditions previously reported to restore the engraftment of cultured cells but did not find that any of these was able to reverse the changes in homing behavior of cytokine-exposed cells. We suggest that these changes in homing and, as a consequence, engraftment result from the increased migratory capacity of infused activated cells, leading to the loss of selectivity of the homing process.

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Adhesion of synchronized human hematopoietic progenitor cells to fibronectin and vascular cell adhesion molecule-1 fluctuates reversibly during cell cycle transit in ex vivo culture

Cited by 21 publications

References 41 publications

The extracellular nucleotide UTP is a potent inducer of hematopoietic stem cell migration

The extracellular nucleotide UTP is a potent inducer of hematopoietic stem cell migration

Functional nanofiber scaffolds with different spacers modulate adhesion and expansion of cryopreserved umbilical cord blood hematopoietic stem/progenitor cells

Impaired bone marrow homing of cytokine-activated CD34+ cells in the NOD/SCID model

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