Differential Kinetics of Primitive Hematopoietic Cells Assayed In Vitro and In Vivo During Serum‐Free Suspension Culture of CD34
            <sup>+</sup>
            Blood Progenitor Cells

Möbest, Dieter; Goan, Silvia‐Renate; Junghahn, I; Winkler, Julia; Fichtner, Iduna; Becker, Michael W.; Lima‐Hahn, Elisete de; Mertelsmann, Roland; Henschler, Reinhard

doi:10.1002/stem.170152

Cited by 39 publications

(25 citation statements)

References 39 publications

(53 reference statements)

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“…BM cells were obtained by flushing the femurs of donor P-selectin-deficient mice in C57BL/6J background (Charles River) as described previously (28). Recipient C57BL/6J mice received lethal irradiation doses at 4 -6 weeks of age, and animals received i.v.…”

Section: Methodsmentioning

confidence: 99%

Endothelial P-Selectin as a Target of Heparin Action in Experimental Melanoma Lung Metastasis

et al. 2004

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Spontaneous and experimental metastasis can be effectively inhibited by the widely used anticoagulant heparin in different tumor models. At the cellular level, many of the antimetastatic effects of heparin in vivo are due to its action on P-selectin-mediated binding. Whereas previous attention has focused on P-selectin-dependent tumor-cell-platelet interactions in blood-borne metastasis, we sought to address the potential contribution of endothelial P-selectin expression to adhesive events between the microvasculature and melanoma cells in vivo. Transplantation of bone marrow from P-selectin-deficient into wild-type mice conveyed inhibition of experimental melanoma metastasis. However, the extent to which bone marrow-conferred lack of platelet P-selectin expression attenuated melanoma lung metastasis was significantly less than that seen in P-selectindeficient mice, suggesting that endothelial P-selectin expression may additionally contribute to formation of hematogenous metastases. This assumption was supported by our intravital microscopy studies, in which a significant proportion of melanoma cells were capable of directly interacting with postcapillary venules of the murine ear in a P-selectin-dependent manner. Heparin not only inhibits P-selectin-mediated melanoma cell rolling but also attenuates melanoma metastasis formation in vivo, further supporting the concept that endothelial P-selectin expression may represent an additional target of heparin action in experimental melanoma lung metastasis.

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Section: Methodsmentioning

confidence: 99%

Endothelial P-Selectin as a Target of Heparin Action in Experimental Melanoma Lung Metastasis

et al. 2004

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“…However, results obtained from stroma-based 1 and stroma-free ex vivo culture systems [2][3][4][5] have been disappointing, owing to insufficient activation of primitive CD34 ϩ CD38 Ϫ cells, cell differentiation, and a loss of repopulating capacity following short-term culture. 6 Moreover, increased CD34 ϩ cell numbers, colony-forming cells (CFCs), and long-term culture initiating cells (LTC-ICs) are not quantitative indicators of in vivo repopulating potential.…”

Section: Introductionmentioning

confidence: 99%

“…37 Of note, both the murine studies and the studies by Gan et al were performed in the absence of exogenous cytokines, which would have been expected to drive differentiation. [4][5][6] In our studies, we supplemented HUBEC monolayers with a cytokine combination, which maximally induced progenitor cell division, and despite this, we observed a measurable increase in SRCs over time. Since exposure to GM-CSF plus IL-3 plus IL-6 plus SCF plus flt-3 ligand in the absence of HUBECs caused a decline in detectable SRCs over 7 days of culture, we postulate that HUBECs may have protected ABM SRCs from differentiation during exposure to cytokines while also supporting the self-renewal of this primitive population.…”

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confidence: 99%

Ex vivo culture with human brain endothelial cells increases the SCID-repopulating capacity of adult human bone marrow

Chute

Saini

Chute

et al. 2002

Blood

104

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Adult human bone marrow (ABM) is an important source of hematopoietic stem cells for transplantation in the treatment of malignant and nonmalignant diseases. However, in contrast to the recent progress that has been achieved with umbilical cord blood, methods to expand ABM stem cells for therapeutic applications have been disappointing. In this study, we describe a novel culture method that uses human brain endothelial cells (HUBECs) and that supports the quantitative expansion of the most primitive measurable cell within the adult bone marrow compartment, the nonobese diabetic/ severe combined immunodeficient (NOD/ SCID) repopulating cell (SRC). Coculture of human ABM CD34 ؉ cells with brain endothelial cells for 7 days supported a 5.4-fold increase in CD34 ؉ cells, induced more than 95% of the CD34 ؉ CD38 ؊ subset to enter cell division, and produced progeny that engrafted NOD/SCID mice at significantly higher rates than fresh ABM CD34 ؉ cells. Using a limiting dilution analysis, we found the frequency of SRCs within fresh ABM CD34 ؉ cells to be 1 in 9.9 ؋ 10 5 cells. Following HUBEC culture, the estimated frequency of SRCs increased to 1 in 2.4 ؋ 10 5 cells. All mice that received transplants of HUBECcultured cells showed B-lymphoid and myeloid differentiation, indicating that a primitive hematopoietic cell was preserved during culture. Noncontact HUBEC cultures also maintained SRCs at a level comparable to contact HUBEC cultures, suggesting that cell-to-cell contact was not required. These data demonstrate that human brain endothelial cells possess a unique hematopoietic activity that increases the repopulating capacity of adult human bone marrow. IntroductionThe development of ex vivo culture methods that promote the expansion of adult human bone marrow (ABM) stem cells would have direct application in clinical gene therapy and stem cell transplantation. However, results obtained from stroma-based 1 and stroma-free ex vivo culture systems [2][3][4][5] have been disappointing, owing to insufficient activation of primitive CD34 ϩ CD38 Ϫ cells, cell differentiation, and a loss of repopulating capacity following short-term culture. 6 Moreover, increased CD34 ϩ cell numbers, colony-forming cells (CFCs), and long-term culture initiating cells (LTC-ICs) are not quantitative indicators of in vivo repopulating potential. [7][8][9][10] Therefore, the importance of evaluating ex vivo cultured cells in an in vivo repopulation model has been emphasized. 8 The nonobese diabetic/severe combined immunodeficient (NOD/ SCID) model system has been used to measure the long-term reconstitution potential of ex vivo-expanded human lymphohematopoietic stem cells. 7-10 SCID-repopulating cells (SRCs) are enriched in human cord blood (CB) as compared with adult ABM and mobilized peripheral blood 11,12 and are most highly concentrated within the CD34 ϩ CD38 Ϫ population. 8 SRCs are considered to be biologically more primitive than assayable LTC-IC and CFC progenitors, 1,9,10,13 which are found in both the CD34 ϩ CD38 ϩ and the CD34 ϩ...

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“…The lack of correlation between SRC and expanded CD34 + cells, CFU and LTC-IC have been demonstrated by various groups. [25][26][27] In spite of the suggestions that human CFU and week-6 CAFC/LTC-IC assays detect highly overlapping populations, 22,23 it remains unclear whether SRC represents an exclusive subset of pluripotent stem cells. Ex vivo expansion of cord blood stem cells has been suggested as a strategy to increase the quantity of progenitor cells required for a successful transplant.…”

Section: Discussionmentioning

confidence: 99%

Cobblestone area-forming cells, long-term culture-initiating cells and NOD/SCID repopulating cells in human neonatal blood: a comparison with umbilical cord blood

Zhang

Fok

et al. 2002

Bone Marrow Transplant

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Differential Kinetics of Primitive Hematopoietic Cells Assayed In Vitro and In Vivo During Serum‐Free Suspension Culture of CD34 ⁺ Blood Progenitor Cells

Abstract: ABSTRACT

Cited by 39 publications

References 39 publications

Endothelial P-Selectin as a Target of Heparin Action in Experimental Melanoma Lung Metastasis

Endothelial P-Selectin as a Target of Heparin Action in Experimental Melanoma Lung Metastasis

Ex vivo culture with human brain endothelial cells increases the SCID-repopulating capacity of adult human bone marrow

Cobblestone area-forming cells, long-term culture-initiating cells and NOD/SCID repopulating cells in human neonatal blood: a comparison with umbilical cord blood

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Differential Kinetics of Primitive Hematopoietic Cells Assayed In Vitro and In Vivo During Serum‐Free Suspension Culture of CD34 + Blood Progenitor Cells

Abstract: ABSTRACT

Cited by 39 publications

References 39 publications

Endothelial P-Selectin as a Target of Heparin Action in Experimental Melanoma Lung Metastasis

Endothelial P-Selectin as a Target of Heparin Action in Experimental Melanoma Lung Metastasis

Ex vivo culture with human brain endothelial cells increases the SCID-repopulating capacity of adult human bone marrow

Cobblestone area-forming cells, long-term culture-initiating cells and NOD/SCID repopulating cells in human neonatal blood: a comparison with umbilical cord blood

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Differential Kinetics of Primitive Hematopoietic Cells Assayed In Vitro and In Vivo During Serum‐Free Suspension Culture of CD34 ⁺ Blood Progenitor Cells