scholarly journals Bone Marrow Mononuclear Cells Activate Angiogenesis via Gap Junction–Mediated Cell-Cell Interaction

Stroke ◽  
2020 ◽  
Vol 51 (4) ◽  
pp. 1279-1289 ◽  
Author(s):  
Akie Kikuchi-Taura ◽  
Yuka Okinaka ◽  
Yukiko Takeuchi ◽  
Yuko Ogawa ◽  
Mitsuyo Maeda ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Gap Junction ◽  
Mononuclear Cells ◽  
Cell Interaction ◽  
Bone Marrow Mononuclear Cells ◽  
Hematopoietic Stem ◽  
Direct Cell ◽  
Cell Cell

Background and Purpose— Bone marrow mononuclear cells (BM-MNCs) are a rich source of hematopoietic stem cells and have been widely used in experimental therapies for patients with ischemic diseases. Activation of angiogenesis is believed to be one of major BM-MNC mode of actions, but the essential mechanism by which BM-MNCs activate angiogenesis have hitherto been elusive. The objective of this study is to reveal the mechanism how BM-MNCs activate angiogenesis. Methods— We have evaluated the effect of direct cell-cell interaction between BM-MNC and endothelial cell on uptake of VEGF (vascular endothelial growth factor) into endothelial cells in vitro. Cerebral ischemia model was used to evaluate the effects of direct cell-cell interaction with transplanted BM-MNC on endothelial cell at ischemic tissue. Results— The uptake of VEGF into endothelial cells was increased by BM-MNC, while being inhibited by blockading the gap junction. Low-molecular-weight substance was transferred from BM-MNC into endothelial cells via gap junctions in vivo, followed by increased expression of hypoxia-inducible factor-1α and suppression of autophagy in endothelial cells. The concentration of glucose in BM-MNC cytoplasm was significantly higher than in endothelial cells, and transfer of glucose homologue from BM-MNC to endothelial cells was observed. Conclusions— Our findings demonstrated cell-cell interaction via gap junction is the prominent pathway for activation of angiogenesis at endothelial cells after ischemia and provided novel paradigm that energy source supply by stem cell to injured cell is one of the therapeutic mechanisms of cell-based therapy. Visual Overview— An online visual overview is available for this article.

scholarly journals Role of microvascular endothelial cells on proliferation, migration and adhesion of hematopoietic stem cells

2020 ◽  
Vol 40 (3) ◽  
Author(s):  
Fanli Lin ◽  
Shuyue Wang ◽  
Hao Xiong ◽  
Yang Liu ◽  
Xiaoming Li ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Endothelial Cells ◽  
Hematopoietic Stem Cells ◽  
Ex Vivo ◽  
Microvascular Endothelial Cells ◽  
Cell Contact ◽  
Hematopoietic Stem ◽  
Direct Cell ◽  
Cell Cell

Abstract Background: The present study investigated the effects of microvascular endothelial cells (MECs) on the chemotaxis, adhesion and proliferation of bone marrow hematopoietic stem cells (HSCs) ex vivo. Methods and Results: MECs were collected from the lung tissue of C57BL/6 mice, and HSCs were isolated with immunomagnetic beads from bone marrow of GFP mice. MECs and HSCs were co-cultured with or without having direct cell–cell contact in Transwell device for the measurement of chemotaxis and adhesion of MECs to HSCs. Experimental results indicate that the penetration rate of HSCs from the Transwell upper chamber to lower chamber in ‘co-culture’ group was significantly higher than that of ‘HSC single culture’ group. Also, the HSCs in co-culture group were all adherent at 24 h, and the co-culture group with direct cell–cell contact had highest proliferation rate. The HSC number was positively correlated with vascular endothelial growth factor (VEGF) and stromal cell-derived factor-1 (SDF-1) levels in supernatants of the culture. Conclusions: Our study reports that MECs enhance the chemotaxis, adhesion and proliferation of HSCs, which might be related to cytokines SDF-1 and VEGF secreted by MECs, and thus MECs enhance the HSC proliferation through cell–cell contact. The present study revealed the effect of MECs on HSCs, and provided a basis and direction for effective expansion of HSCs ex vivo.

Enrichment of CD34+ Hematopoietic Stem and Progenitor Cells from Human Bone Marrow Using a P-Selectin-Coated Microtube.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 1219-1219
Author(s):  
Srinivas D. Narasipura ◽  
Jane L. Liesveld ◽  
Joel C. Wojciechowski ◽  
Nichola Charles ◽  
Karen Rosell ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Progenitor Cells ◽  
Adhesion Molecule ◽  
Mononuclear Cells ◽  
Single Cells ◽  
Bone Marrow Mononuclear Cells ◽  
Cell Capture ◽  
Hematopoietic Stem ◽  
Stem And Progenitor Cells ◽  
Coated Surfaces

Abstract Enrichment and purification of hematopoietic stem and progenitor cells (HSPCs) is important in transplantation therapies for hematological disorders and for basic stem cell research. Primitive CD34+ HSPCs have demonstrated stronger rolling adhesion than mature CD34- mononuclear cells on selectins (Blood2000; 95:478–486). We have exploited this differential rolling behavior to capture and purify HSPCs from bone marrow, by perfusing mononuclear cells through selectin-coated microtubes. Bone marrow mononuclear cells were perfused through the cell capture microtubes coated with adhesion molecules. These utilized a parallel plate flow chamber (Glycotech), and the P-selectin was adsorbed with laboratory tubing of appropriate lengths attached to the inlet, outlet, and vacuum ports of the gasket chamber. After perfusion, the device lumen was washed and captured cells were visualized and estimated by video microscopy. “Rolling” cells were defined as cells translating at less than 50% of the calculated hydrodynamic free stress velocity. Velocities of single cells were determined using a MATLAB program designed to measure the change in position of the cell centroid in a given time period. Adherent cells were eluted by high shear, calcium free buffer and air embolism. Immunofluorescence staining followed by flow cytometry was used to analyze CD34+ HSPCs. CD34+ HSPC purity of cells captured in adhesion molecule-coated devices was significantly higher than the fraction of CD34+ cells found in bone marrow- mononuclear cells (2.5 ± 0.8%). P-selectin coated surfaces yielded 16–20% CD34+ cell purity, while antibody coated surfaces yielded 12–18%. Although the CD34+ cell purities were comparable between selectin and antibody surfaces, the total number of CD34+ HSPCs captured was significantly higher in P-selectin devices (∼5.7–7.1 × 104) when compared to the antibody device (∼1.74–2.61 × 104). Furthermore, analysis for cells positive for CD133, a surface marker for more primitive HSPCs, depicted approximately 10–14 fold enrichment in P-selectin samples over control bone marrow mononuclear cells. The captured cells were viable and exhibited in vitro colony forming capabilities. Thus, P-selectin can be used in a compact flow device to capture and enrich HSPCs. This study supports the hypothesis that flow-based adhesion molecule-mediated capture may be a viable physiologic approach to the capture and purification of HSPCs.

scholarly journals Neuropilin 1 regulates bone marrow vascular regeneration and hematopoietic reconstitution

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Christina M. Termini ◽  
Amara Pang ◽  
Tiancheng Fang ◽  
Martina Roos ◽  
Vivian Y. Chang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Hematopoietic Stem Cells ◽  
Hematopoietic Stem ◽  
Vascular Regeneration ◽  
Vascular Niche ◽  
Hematopoietic Reconstitution ◽  
Neuropilin 1

AbstractIonizing radiation and chemotherapy deplete hematopoietic stem cells and damage the vascular niche wherein hematopoietic stem cells reside. Hematopoietic stem cell regeneration requires signaling from an intact bone marrow (BM) vascular niche, but the mechanisms that control BM vascular niche regeneration are poorly understood. We report that BM vascular endothelial cells secrete semaphorin 3 A (SEMA3A) in response to myeloablation and SEMA3A induces p53 – mediated apoptosis in BM endothelial cells via signaling through its receptor, Neuropilin 1 (NRP1), and activation of cyclin dependent kinase 5. Endothelial cell – specific deletion of Nrp1 or Sema3a or administration of anti-NRP1 antibody suppresses BM endothelial cell apoptosis, accelerates BM vascular regeneration and concordantly drives hematopoietic reconstitution in irradiated mice. In response to NRP1 inhibition, BM endothelial cells increase expression and secretion of the Wnt signal amplifying protein, R spondin 2. Systemic administration of anti - R spondin 2 blocks HSC regeneration and hematopoietic reconstitution which otherwise occurrs in response to NRP1 inhibition. SEMA3A – NRP1 signaling promotes BM vascular regression following myelosuppression and therapeutic blockade of SEMA3A – NRP1 signaling in BM endothelial cells accelerates vascular and hematopoietic regeneration in vivo.

Primary endothelial cells isolated from the yolk sac and para-aortic splanchnopleura support the expansion of adult marrow stem cells in vitro

Blood ◽  
2003 ◽  
Vol 102 (13) ◽  
pp. 4345-4353 ◽  
Author(s):  
Weiming Li ◽  
Scott A. Johnson ◽  
William C. Shelley ◽  
Michael Ferkowicz ◽  
Paul Morrison ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Ex Vivo ◽  
Yolk Sac ◽  
Hematopoietic Progenitor Cell ◽  
Hematopoietic Progenitor ◽  
Hematopoietic Stem

AbstractThe embryonic origin and development of hematopoietic and endothelial cells is highly interdependent. We hypothesized that primary endothelial cells from murine yolk sac and para-aortic splanchnopleura (P-Sp) may possess the capacity to expand hematopoietic stem cells (HSCs) and progenitor cells ex vivo. Using Tie2-GFP transgenic mice in combination with fluorochrome-conjugated monoclonal antibodies to vascular endothelial growth factor receptor-2 (Flk1) and CD41, we have successfully isolated pure populations of primary endothelial cells from 9.5-days after coitus (dpc) yolk sac and P-Sp. Adult murine bone marrow Sca-1+c-Kit+lin- cells were cocultured with yolk sac or P-Sp Tie2-GFP+Flk-1+CD41- endothelial cell monolayers for 7 days and the total number of nonadherent cells increased 47- and 295-fold, respectively, and hematopoietic progenitor counts increased 9.4- and 11.4-fold, respectively. Both the yolk sac and P-Sp endothelial cell cocultures facilitated long-term (> 6 months) HSC competitive repopulating ability (2.8- to 9.8-fold increases, respectively). These data suggest that 9.5-dpc yolk sac- and P-Sp-derived primary Tie2-GFP+Flk-1+CD41- endothelial cells possess the capacity to expand adult bone marrow hematopoietic progenitor cell and HSC repopulating ability ex vivo. (Blood. 2003;102:4345-4353)

N-Cadherin and Cadherin-11 Play Vital Roles in the Cell-Cell Contact between Hematopoietic Progenitor Cells and Mesenchymal Stromal Cells.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 1406-1406
Author(s):  
Wolfgang Wagner ◽  
Frederik Wein ◽  
Anke Diehlmann ◽  
Rainer Saffrich ◽  
Patrick Wuchter ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stromal Cells ◽  
Cell Interaction ◽  
Hematopoietic Progenitor Cells ◽  
Feeder Layer ◽  
Primitive Function ◽  
Hematopoietic Progenitor ◽  
Cd34 Cells ◽  
Direct Cell ◽  
Cell Cell

Abstract Self renewal and differentiation of hematopoietic progenitor cells (HPC) are regulated by the microenvironment of the bone marrow. As an in vitro model system, mesenchymal stromal cells (MSC) provide a supportive cellular microenvironment for maintaining primitive function of HPC. It has been postulated that direct cell-cell interaction is crucial for maintenance of “stemness”. Human HPC were co-cultured with MSC from human bone marrow and subsequently separated into an adherent and a non-adherent fraction. HPC subsets with higher self-renewing capacity demonstrated significantly higher adhesion to MSC (CD34+vs. CD34−, CD34+/CD38−vs. CD34+/CD38+, slow dividing fraction vs. fast dividing fraction). Long-term culture-initiating cell (LTC-IC) frequency was higher in the adherent fraction than in the non-adherent fraction of CD34+ cells. Microarray analysis (Affymetrix, U133_Plus_2.0) revealed that differentially expressed genes coding for adhesion proteins were highly up-regulated in the adherent fraction of CD34+ cells. These genes included VCAM1, connexin 43 and cadherin-11. Furthermore, we have compared the supportive potential of different feeder layer preparations. Human MSC were isolated from bone marrow (BM), from adipose tissue (AT) and umbilical cord blood (CB). The ability to maintain LTC-IC and a primitive CD34+CD38− immunophenotype was significantly higher for MSC derived from BM and CB compared to those from AT. These results were in line with higher adhesion of HPC to BM-MSC and CB-MSC in comparison to AT-MSC. Analysis of the cytokine production of MSC preparations by antibody arrays, ELISA and by a cytometric bead array showed that albeit there were significant differences in the chemokine secretion profiles of the aforementioned MSC preparations, there was no relationship to their potentials in maintaining primitive function of HPC. Global gene expression profiles of MSC preparations showed that adhesion proteins including N-cadherin, cadherin-11, VCAM1, NCAM1 and integrins were highly expressed in MSC preparations derived from BM and CB. Western blot analysis confirmed higher protein expression of N-cadherin and cadherin-11 in BM-MSC compared to AT-MSC and CB-MSC. Fluorescent microscopic analysis revealed that N-cadherin is located at the cell-cell contacts between HPC and MSC. Expression of N-cadherin or cadherin-11 was efficiently knocked down in MSC feeder layer using siRNA. This effect was verified by Western blot analysis and it lasted for up to seven days. Adhesion of HPC was significantly reduced on MSC that have been treated by siRNAs for N-cadherin and cadherin-11 whereas siRNA for MAPK did not affect cell-cell interaction. Similarly, a blocking functional antibody for N-cadherin reduced significantly the adhesion of HPC to MSC. MSC provide a microenvironment which supports the maintenance of primitive function of HPC. Our results indicated that direct cell-cell interaction mediated by N-cadherin and cadherin-11 plays a central role in this interaction of HPC with their cellular microenvironment.

Targeting Leukemia Initiating Cells by MUC1-C Subunit Inhibition

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 3583-3583
Author(s):  
Dina Stroopinsky ◽  
Jacalyn Rosenblatt ◽  
Heidi Mills ◽  
Caterina Nardella ◽  
Keisuke Ito ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Cell Morphology ◽  
Mononuclear Cells ◽  
Leukemia Cells ◽  
Bone Marrow Mononuclear Cells ◽  
Hematopoietic Stem ◽  
Nsg Mice ◽  
C Subunit ◽  
Normal Human

Abstract Abstract 3583 Introduction: Acute myeloid leukemia (AML) arises from a population of leukemia initiating cells (LICs) that exhibit chemotherapy resistance and serve as a reservoir for disease recurrence. We have demonstrated that the oncoprotein, MUC1, is expressed on LICs but not normal hematopoietic stem cells. We have developed an inhibitor of the MUC1-C subunit, designated GO-203, which blocks MUC1-C dimerization and its oncogenic function. In a murine NSG (NOD-scid IL2Rg) model, we examined whether MUC1high progenitors are more effective in inducing leukemic engraftment as compared to MUC1low progenitors and evaluated the capacity of GO-203 to eradicate disease. Methods and results: To confirm the hypothesis that LICs are contained within the MUChigh progenitor population, lineage−/MUC1high and lineage−/MUC1low cells were isolated from AML derived bone marrow mononuclear cells by flow cytometric sorting, and transplanted (1×106 cells/mouse) into sub-lethally irradiated NSG mice using retro orbital injections. Leukemia engraftment was observed in all recipient mice (6/6) inoculated with MUC1high cells, with no evidence of normal human engraftment as shown by immunophenotype, cell morphology, and presence of cytogenetic abnormality. In a cohort of 6 mice, treatment with GO-203 was initiated 8 weeks after challenge with MUC1high AML cells following the demonstration of circulating leukemia cells. Leukemia was eradicated in 5/6 mice treated with GO-203. In contrast, 6/6 mice inoculated with MUC1low cells demonstrated engraftment with normal human hematopoetic cells as demonstrated by immunophenotype, cell morphology and absence of cytogenetic abnormality by FISH analysis. Engraftment of normal hematopoietic cells was unaffected by treatment with GO-203 in 4/5 of mice inoculated with MUC1low cells, supporting that GO-203 does not affect non-malignant hematopoiesis. We further determined the effect of GO-203 treatment in a prevention model. NSG mice were treated with 21 days of GO-203 beginning 24 hours after inoculation with CD34+/lineage−/MUC1high cells isolated from patient derived bone marrow mononuclear cells. Treatment with GO-203 prevented leukemia engraftment in 4/5 animals with the 5th animal showing only 1.9% leukemia cells. In contrast, 4/5 animals inoculated with the CD34+/lineage-/MUC1high cells in the absence of GO-203 demonstrated evidence of extensive bone marrow infiltration with leukemia cells comprising a mean of 33% of the bone marrow mononuclear cells. Conclusions: The results demonstrate that LICs are contained within the MUC1high fraction of AML derived progenitors and that MUC1low progenitors contain normal hematopoietic stem cells capable of normal engraftment. Treatment with the MUC1-C inhibitor, GO-203, prevents engraftment of leukemia, and eradicates established disease in a murine model. Our work indicates that GO-203 is a novel therapeutic agent for AML that targets LICs and has the potential to improve disease outcomes. Disclosures: Kufe: Genus Oncology: Consultancy.

scholarly journals Direct cell-cell interaction of cardiomyocytes is key for bone marrow stromal cells to go into cardiac lineage in vitro

2003 ◽  
Vol 125 (6) ◽  
pp. 1470-1479 ◽  
Author(s):  
Shinya Fukuhara ◽  
Shinji Tomita ◽  
Seiji Yamashiro ◽  
Takayuki Morisaki ◽  
Chikao Yutani ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stromal Cells ◽  
Bone Marrow Stromal Cells ◽  
Cell Interaction ◽  
Marrow Stromal Cells ◽  
Direct Cell ◽  
Cardiac Lineage ◽  
Cell Cell

Transplanted adult hematopoietic stems cells differentiate into functional endothelial cells

Blood ◽  
2004 ◽  
Vol 103 (1) ◽  
pp. 13-19 ◽  
Author(s):  
Alexis S. Bailey ◽  
Shuguang Jiang ◽  
Michael Afentoulis ◽  
Christina I. Baumann ◽  
David A. Schroeder ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Hematopoietic Stem Cells ◽  
Blood Vessels ◽  
Cell Potential ◽  
Hematopoietic Stem ◽  
Adult Bone Marrow ◽  
Adult Bone

Abstract During early embryogenesis, blood vessels and hematopoietic cells arise from a common precursor cell, the hemangioblast. Recent studies have identified endothelial progenitor cells in the peripheral blood, and there is accumulating evidence that a subset of these cells is derived from precursors in the bone marrow. Here we show that adult bone marrow–derived, phenotypically defined hematopoietic stem cells (c-kit+, Sca-1+, lineage–) give rise to functional endothelial cells. With the exception of the brain, donor-derived cells are rapidly integrated into blood vessels. Durably engrafted endothelial cells express CD31, produce von Willebrand factor, and take up low-density lipoprotein. Analysis of DNA content indicates that donor-derived endothelial cells are not the products of cell fusion. Self-renewal of stem cells with hematopoietic and endothelial cell potential was revealed by serial transplantation studies. The clonal origin of both hematopoietic and endothelial cell outcomes was established by the transfer of a single cell. These results suggest that adult bone marrow–derived hematopoietic stem cells may serve as a reservoir for endothelial cell progenitors.

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