scholarly journals Hemogenic Endothelium-like Changes Were Derived to an Adult Human Dermal Fibroblast By Electrical Stimulation

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 5129-5129
Author(s):  
Takuji Matsuo ◽  
Ryosuke Shirasaki ◽  
Oka Yoko ◽  
Tadashi Yamamoto ◽  
Jun Ooi ◽  
...  
Keyword(s):  
Stem Cell ◽  
Electrical Stimulation ◽  
Endothelial Cell ◽  
Cell Fate ◽  
Dermal Fibroblast ◽  
Vascular Endothelial Cell ◽  
Human Dermal Fibroblast ◽  
Vascular Endothelial ◽  
Hematopoietic Stem ◽  
Morphologic Changes

Abstract Background and Aims: We previously reported that when an adult human dermal fibroblast (HDF) was cultured with interleukin (IL)-1-beta (b) and erythropoietin (EPO), hematopoiesis-related molecules expressed. And, lymphatic duct-neogenesis genes also expressed, and vascular endothelial growth factor (VEGF)-A, and -C were produced. When anti-human VEGF-C antibody (Ab) was added to the cultures, hematopoiesis-related genes expressed; however, morphologic changes were not observed (54th ASH, 18th EHA). Reported findings on nuclear transfer reveal that electrical stimulation induces nuclear fusion and changes cell-fate. We observed the effect of electrical stimulation to IL-1-b-stimulated HDF. Materials and Methods: HDF was cultured with IL-1-b, EPO, VEGF-A, and anti-human VEGF-C Ab for 14 days. Then, cells were suspended in electrical stimulation-buffer (0.25 M d-sorbitol, 0.1 mM Ca-acetate, 0.5 mM Mg-acetate, 1 mg/mL fatty-acid-free BSA, 0.5 mM HEPES), and incubated on ice for 10 minutes. Cells were stimulated at 110V 20mA (0.2 cm electrode, gap), and were further cultured with k/o DMEM containing 20% KSR and SCF, IL-6, FL, IGF-2, and VEGF-A. Morphological changes and expressions of vascular endothelial cell-related genes, and hematopoiesis-related ones were observed. Results: When HDF was stimulated electrically, a few cells showed vascular endothelial cell-morphology after two days. When cells were further cultured in a hematopoietic stem cell-culturing condition, a part of endothelial cell-morphology changed to that of non-adherent hematopoietic cells or blastic colonies, in which increased expression-levels of SCL, CD41, GATA-2, CD34, and CD45 were accepted with significant statistical difference. Discussion: Recent reports reveal that a kind of vascular endothelial cells, called hemogenic endothelium, can convert to a hematopoietic cell, and works an important role in hematopoietic stem cell-generation. We hypothesized that when cells expressed a kind of specific transcripts, some stimulation triggerred to convert morphologic changes and cell-fate as are observed in nuclear fusion. And, our observations indicated that when IL-1-b-stimulated HDF, expressing hematopoiesis-related transcripts, were stimulated electrically, cellular morphology changed. Currently, we are precisely analyzing electrically stimulated HDF’s biological characteristics. Disclosures No relevant conflicts of interest to declare.

Platelet Transfusion Requirements Are Associated with Endothelial Cell Injury and Angiogenesis During Allogeneic Hematopoietic Stem Cell Transplantation.

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 3487-3487
Author(s):  
Marianne E. McPherson ◽  
Benjamin Yaw Owusu ◽  
Keith J. August ◽  
Muna Qayed ◽  
Robert H. Lyles ◽  
...  
Keyword(s):  
Stem Cell ◽  
Endothelial Cell ◽  
Growth Factor ◽  
Linear Regression ◽  
Poor Response ◽  
Vascular Endothelial ◽  
Simple Linear Regression ◽  
Hematopoietic Stem ◽  
Relationship Of ◽  
The Relationship

Abstract Abstract 3487 Hematopoietic stem cell transplantation (HSCT) is associated with poor response to platelet (PLT) transfusions. It has been posited that vascular endothelial cell (EC) damage from the effects of high dose chemoradiotherapy is responsible for PLT consumption. To test this hypothesis, we assessed the relationship between serial levels of plasma markers of vascular damage and repair, and PLT transfusion requirement. Plasma samples were obtained on 40 adult and pediatric recipients of allogeneic HSCT with myeolablative conditioning for leukemia, lymphoma, or myelodysplastic syndrome at days -10, +5, +15, and +30. Single donor apheresis PLT transfusions were given prophylactically for PLT count <10,000/mcL or as needed for hemorrhage. To minimize the influence of the rate of engraftment only transfusions through day +15 were considered. The number of transfused PLT units divided by patient's body weight (BW) was recorded. For patients with BW >125% of ideal BW, adjusted BW was calculated as 0.25 × (actual – ideal BW) + ideal BW. The influence of response to PLT transfusions prior to transplant admission was considered. Poor response to PLT transfusion was defined as a PLT count rise of <30,000/mcL on the last PLT transfusion prior to HSCT; individuals with poor response to PLT transfusions prior to HSCT were not excluded. Plasma levels of angiopoietin-2 (Ang-2), platelet-derived growth factor-BB (PDGF-BB), soluble Platelet Endothelial Cell Adhesion Molecule (sPECAM), and vascular endothelial growth factor (VEGF) at each time point were measured using a multiplex ELISA, and the ratio of each biomarker level at days +5 +15, and +30 to the baseline level (day -10 pre-HSCT) was calculated. The ratios of the biomarker level at each time point to baseline were compared by 1-way ANOVA. The relationship of cumulative PLT transfusions at day +15 to the logarithm of each biomarker ratio was assessed by Pearson's correlation coefficient. Simple linear regression was used to assess the relationships of age, gender, total body irradiation (TBI), donor matching, stem cell source, hemorrhage, hepatic veno-occlusive disease (VOD), and prior response to PLTs with cumulative PLT transfusions, and significant variables were introduced into multivariable analysis to assess the relationship of significant biomarker ratios to PLT transfusions. Four patients were excluded from analysis, 1 for death at day +13 and 3 for insufficient samples. The median age was 17.0 years (range 6.7–55.4 years). Nine patients received HLA matched related donor transplants and 31 received alternative donor transplants. TBI was used as part of conditioning in 20 patients, VOD developed in 5, and hemorrhage occurred in 13. The mean number of platelet transfusions from days -10 – 15 was 1.1 units/10 kg BW (range 0.12–3.7 units/10 kg). Ten patients had poor responses to PLT prior to HSCT, 21 had good responses to PLT, and 5 had insufficient data regarding PLT response. Significant reductions were observed in sPECAM-1 at day 5 (mean ratio 0.58 of baseline, range 0.29–1.05, p<0.0001) and in PDGF-BB (mean ratio 0.59 of baseline, range 0.03–2.44, p=0.0198). VEGF was mildly increased over baseline at day 5 (mean ratio 1.49 of baseline, range 0.29–8.4, p=0.048). Ang-2 level was increased over baseline at days 5 and 15, peaking at 8.8-times greater than baseline on day 15 (p=0.075). The log of (Ang-2 day 15/Ang-2 baseline) had a significant positive correlation with cumulative PLT transfusions at day 15 (r2=0.32, p=0.05) (Figure). In simple linear regression, PLT transfusion at day 15 also was associated with hemorrhage (p=0.008) and VOD (p=0.083). Poor PLT response prior to HSCT was not associated with cumulative PLT transfusions at day 15 (p=0.47). In multiple linear regression controlling for hemorrhage and VOD, the association of log(Ang-2 day 15/Ang-2 baseline) with cumulative PLT transfusions remained significant (p=0.024). In conclusion, significant decreases in plasma levels of the vascular adhesion molecule sPECAM-1 and angiogenic growth factor PDGF-BB are seen at day +5 in HSCT. There was wide variation in Ang-2 levels; however increased Ang-2 at day +15 was associated with increased cumulative PLT transfusions at day +15, suggesting that vascular regeneration is associated with increased PLT consumption. Disclosures: Ofori-Acquah: Emory University: Patents & Royalties.

scholarly journals Driving vascular endothelial cell fate of human multipotent Isl1+ heart progenitors with VEGF modified mRNA

Cell Research ◽  
2013 ◽  
Vol 23 (10) ◽  
pp. 1172-1186 ◽  
Author(s):  
Kathy O Lui ◽  
Lior Zangi ◽  
Eduardo A Silva ◽  
Lei Bu ◽  
Makoto Sahara ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Cell Fate ◽  
Vascular Endothelial Cell ◽  
Vascular Endothelial

scholarly journals Acute and endothelial-specific Robo4 deletion affect hematopoietic stem cell trafficking independent of VCAM1

PLoS ONE ◽  
2021 ◽  
Vol 16 (8) ◽  
pp. e0255606
Author(s):  
Stephanie Smith-Berdan ◽  
Alyssa Bercasio ◽  
Leah Kramer ◽  
Bryan Petkus ◽  
Lindsay Hinck ◽  
...  
Keyword(s):  
Stem Cell ◽  
Endothelial Cell ◽  
Hematopoietic Stem Cell ◽  
Transmembrane Protein ◽  
Vascular Endothelial Cell ◽  
Surface Protein ◽  
Cell Trafficking ◽  
Wild Type ◽  
Normal Numbers ◽  
Hematopoietic Stem

Hematopoietic stem cell (HSC) trafficking is regulated by a number of complex mechanisms. Among them are the transmembrane protein Robo4 and the vascular cell adhesion molecule, VCAM1. Endothelial VCAM1 is a well-known regulator of hematopoietic cell trafficking, and our previous studies revealed that germline deletion of Robo4 led to impaired HSC trafficking, with an increase in vascular endothelial cell (VEC) numbers and downregulation of VCAM1 protein on sinusoidal VECs. Here, we utilized two Robo4 conditional deletion models in parallel with Robo4 germline knockout mice (R4KO) to evaluate the effects of acute and endothelial cell-specific Robo4 deletion on HSC trafficking. Strikingly similar to the R4KO, the acute deletion of Robo4 resulted in altered HSC distribution between the bone marrow and blood compartments, despite normal numbers of VECs and wild-type levels of VCAM1 cell surface protein on sinusoidal VECs. Additionally, consistent with the R4KO mice, acute loss of Robo4 in the host perturbed long-term engraftment of donor wild-type HSCs and improved HSC mobilization to the peripheral blood. These data demonstrate the significant role that endothelial Robo4 plays in directional HSC trafficking, independent of alterations in VEC numbers and VCAM1 expression.

scholarly journals Effects of Coculture Fibroblasts and Vascular Endothelial Cells on Proliferation and Osteogenesis of Adipose Stem Cells

2022 ◽  
Vol 2022 ◽  
pp. 1-11
Author(s):  
Xinyu Liao ◽  
Ruiying Zhong ◽  
Hong Zhang ◽  
Fuke Wang
Keyword(s):  
Stem Cells ◽  
Endothelial Cells ◽  
Stem Cell ◽  
Endothelial Cell ◽  
Vascular Endothelial Cells ◽  
Vascular Endothelial Cell ◽  
Adipose Stem Cells ◽  
Cell Group ◽  
Vascular Endothelial ◽  
Adipose Stem Cell

Background. The development of tissue engineering provides a new method for the clinical treatment of bone defects, but the problems of slow formation and slow vascularization of tissue engineered bone have always existed. Studies have shown that the combined culture system of vascular endothelial cells and adipose stem cells is superior to single cell in repairing bone defects. With the excellent proliferation ability, secretion of synthetic collagen and a variety of regulatory factors and fibroblasts can differentiate into osteoblasts and have the potential to be excellent seed cells involved in tissue engineering bone construction. Objective. To investigate the effects of combined culture of fibroblasts, vascular endothelial cells, and adipose stem cells on proliferation and osteogenic differentiation of adipose stem cells. Methods. The cells were divided into 4 groups: adipose stem cell group, adipose stem cell+vascular endothelial cell coculture group, adipose stem cell+fibroblast coculture group, and adipose stem cell+vascular endothelial cell+fibroblast coculture group. The morphological changes of the cells were observed under an inverted microscope. After 1, 3, 5, 7, and 9 days of coculture, the proliferation of adipose stem cells in each group was detected by a CCK-8 method and the growth curve was plotted. Adipose stem cells in each group were stained with alizarin red and alkaline phosphatase at days 7, 14, 21, and 28. At the third week of coculture, Western blot was used to detect the expression level of bone morphogenetic protein 2 of adipose stem cells in each group. Results and Conclusions. (1) After 14 days of culture, some cells in the adipose stem cell+vascular endothelial cell+fibroblast coculture group fused into clumps and distributed in nests, while the adipose stem cells in the adipose stem cell group had a single cell morphology and no cell clusters were observed. (2) The cell growth curves were basically the same in each group, and the absorbance value increased gradually. The absorbance value of the adipocyte+vascular endothelial cell+fibroblast coculture group was the highest, followed by the adipocyte+fibroblast coculture group and then the adipocyte+fibroblast coculture group. (3) Alizarin red staining showed negative reaction in each group on the 7th day, and a small number of red positive cells gradually appeared in each group as time went on. On the 28th day, red positive cells were found in all groups, and most of them were in the coculture group of adipose stem cells+vascular endothelial cells+fibroblasts, showing red focal. The coculture group of adipose stem cells+vascular endothelial cells and adipose stem cells+fibroblasts was less, and the adipose stem cell group was the least. On day 28 of alkaline phosphatase staining, cells in each group had red positive particles, and the adipose stem cell+vascular endothelial cell+fibroblast coculture group and adipose stem cell+fibroblast coculture group had the most, followed by the adipose stem cell+vascular endothelial cell coculture group and then the adipose stem cell group. (4) Bone morphogenetic protein 2 was expressed in all groups, especially in adipose stem cell+fibroblast coculture group and adipose stem cell+vascular endothelial cell+ fibroblast coculture group. (5) Fibroblast could promote adipose stem cell osteogenic differentiation better than vascular endothelial cells, but the proliferation effect was not as good as vascular endothelial cells. The coculture system of fibroblast combined with vascular endothelial cells and adipose stem cells promoted the proliferation of adipose stem cells and the rapid and efficient differentiation of adipose stem cells into osteoblasts.

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