Electron microscopy of endothelial cells in culture: I. Transmission electron microscopy

1986 ◽  
Vol 10 (1) ◽  
pp. 31-33 ◽  
Author(s):  
U. S. Ryan ◽  
M. A. Hart
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Endothelial Cells ◽  
Cells In Culture ◽  
Transmission Electron

Secretion of Weibel-Palade bodies observed in extra-alveolar vessels of rabbit lung

1983 ◽  
Vol 54 (5) ◽  
pp. 1284-1286 ◽  
Author(s):  
J. M. McNiff ◽  
J. Gil
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Endothelial Cells ◽  
Pulmonary Artery ◽  
Rabbit Lung ◽  
Vascular Lumen ◽  
Cells In Culture ◽  
Transmission Electron ◽  
Specific Granules

We fixed rabbit lungs by perfusion of osmium into the pulmonary artery and examined in light and transmission electron microscopy a large number of extra-alveolar vessels with a diameter of 0.1–0.25 mm, with emphasis on Weibel-Palade bodies (endothelial specific granules). Weibel-Palade bodies are organelles specific to endothelial cells. Their function is unknown, but they are useful markers for identification of endothelial cells in culture. We were able to observe release of the content of these bodies into the vascular lumen; this indicates that they are secretory.

Histochemical Studies On The In Vitro 5-HT Uptake In Endothelial Cells

1981 ◽  
Author(s):  
T Kjellström ◽  
H Ahlman ◽  
F Dahlström ◽  
G Hansson ◽  
B Stenberg ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Endothelial Cells ◽  
Factor Viii ◽  
Serotonin Uptake ◽  
Adult Rats ◽  
Pulmonary Endothelial Cells ◽  
Transmission Electron ◽  
5 Ht Uptake

Previous studies have shown that 5-HT is rapidly taken up by the endothelial cells and some investigations also suggested that serotonin is metabolized within these cells. In earlier studies on rat-lungs using a fluorescence histo- chemical method according to Hillarp - Falk we demonstrated that 5-HT was accumulated within the mast cells. Using this technique we could not demonstrate any specific uptake in the pulmonary endothelial cells. It was the purpose of the present investigation to further study the 5-HT uptake by isolated pulmonary endothelial cells.Methods Cells from the vascular intima of the pulmonary artery in adult rats were grown in a growth medium containing FCS. The endothelial nature of these cells was demonstrated using transmission electron microscopy and factor VIII analysis. Confluent endothelial cells were incubated with 5-HT and the cellular uptake was studied with fluorescence microscopy according to the Hillarp - Falk procedure.Results The endothelial cells were identified by the presence of Weibel-Palade bodies using transmission electron microscopy and the immunofluorescent demonstration of cellular factor VIII antigen. Cells not exposed to serotonin had no specific 5-HT fluorescense. After incubation with 5-HT at different concentrations there was a progressive uptake of the amine within the cells.Conclusions This study confirms previous reports on the specific serotonin uptake in endothelial cells. The Hillarp-Falk procedure seems suitable for further studies of serotonin uptake in cultured endothelial cells.

Oxyhemoglobin-induced apoptosis in cultured endothelial cells

1999 ◽  
Vol 91 (3) ◽  
pp. 459-465 ◽  
Author(s):  
Kotaro Ogihara ◽  
Alexander Y. Zubkov ◽  
David H. Bernanke ◽  
Adam I. Lewis ◽  
Andrew D. Parent ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Endothelial Cells ◽  
Time Dependent ◽  
Dependent Manner ◽  
Content Type ◽  
Aortic Endothelial Cells ◽  
Bovine Aortic Endothelial Cells ◽  
Transmission Electron ◽  
Fine Print

Object. Oxyhemoglobin (OxyHb) is one of the most important spasmogens for cerebral vasospasm that follows aneurysmal subarachnoid hemorrhage. The cytotoxic effect of OxyHb has been documented in endothelial and smooth-muscle cells; however, the pattern of cell death—necrosis or apoptosis—as the final stage of cell damage has not been demonstrated. This study was undertaken to determine if OxyHb induces apoptotic changes in cultured bovine aortic endothelial cells.Methods. Confluent bovine aortic endothelial cells were treated with OxyHb in a concentration- and time-dependent manner. Cell density was assayed by counting the number of cells that attached to culture dishes after exposure to OxyHb. To identify apoptotic changes, the investigators used three specific methods: DNA fragmentation (electrophoreses), the apoptotic body (transmission electron microscopy), and cleavage of poly (adenosine diphosphate ribose) polymerase (PARP [Western blotting]).Conclusions. Oxyhemoglobin decreased cell density in a concentration- and time-dependent manner. Analysis of DNA showed a pattern of internucleosomal cleavage characteristic of apoptosis (DNA ladder). Transmission electron microscopy demonstrated condensation of nuclei and apoptotic bodies in OxyHb-treated endothelial cells. Western blotting with the PARP antibody revealed that the 116-kD PARP was cleaved to the 85-kD apoptosis-related fragment. These results for the first time demonstrated that the OxyHb induces apoptosis in cultured endothelial cells.

scholarly journals Inhibition of liver sinusoidal endothelial cells in rats with hepatic fibrosis by injection of VEGF 165 via the inferior vena cava

2021 ◽  
Author(s):  
Zhigang Sun ◽  
Tianyi Dong ◽  
Zhun Zhang ◽  
Tiantian Wang ◽  
Chenyu Zhang ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Endothelial Cells ◽  
Liver Fibrosis ◽  
Serum Level ◽  
Basement Membranes ◽  
Liver Sinusoidal Endothelial Cells ◽  
Sinusoidal Endothelial Cells ◽  
Transmission Electron

Abstract Background Although VEGF can maintain the normal phenotype of liver sinusoidal endothelial cells (LSECs), it has also been reported that VEGF exacerbates cirrhosis. The role of VEGF in the progression and recovery of cirrhosis has still remained controversial.Methods We established a cirrhotic rat model by thioacetamide that was used as drinking water; besides, 0, 1, 2, and 4 μg VEGF165 were then continuously injected into the rats. The serum level of hyaluronic acid was measured by ELISA at 0, 1, and 4 weeks, separately. Serum levels of ALT, AST, direct bilirubin, indirect bilirubin, and ALB were detected by an automatic biochemical analyzer. In addition, the levels of VEGF165, CD44, MMP9, MMP2, HIF-1α, and endothelin were detected by Western blotting. The expression level of CD44 in LSECs was detected by immunohistochemistry. Changes of fenestrations of LSECs and basement membranes of blood vessels were observed by transmission electron microscopy. Results With the increase of dosage and duration of VEGF treatment, the levels of liver function markers in the serum, the levels of CD44, HIF-1α, hydroxyproline and endothelin were significantly improved. With determination of the serum level of hydroxyproline in the blood, it was disclosed that the mentioned level was markedly decreased. In the Sirius Red staining, the stained red area was gradually reduced. Images captured by transmission electron microscopy also confirmed that the ultrastructure of LSECs tended to be normal.Conclusion VEGF165 can accelerate the resolution of liver fibrosis by promoting fenestration structure formation in LSECs, as well as promoting material exchange between sinusoids and hepatocytes. Our findings may provide a new insight for the study of the role of VEGF in liver fibrosis.

scholarly journals Mesenchymal stem cell-derived exosomes inhibit the VEGF-A expression in human retinal vascular endothelial cells induced by high glucose

2021 ◽  
Vol 14 (12) ◽  
pp. 1820-1827
Author(s):  
Guang-Hui He ◽  
◽  
Meng Dong ◽  
Song Chen ◽  
Yu-Chuan Wang ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Endothelial Cells ◽  
High Glucose ◽  
Vascular Endothelial Cells ◽  
Vascular Endothelial ◽  
Nanoparticle Tracking Analysis ◽  
Transmission Electron ◽  
Group B ◽  
Group D

AIM: To determine the effect of exosomes derived from human umbilical cord blood mesenchymal stem cells (hUCMSCs) on the expression of vascular endothelial growth factor A (VEGF-A) in human retinal vascular endothelial cells (HRECs). METHODS: Exosomes were isolated from hUCMSCs using cryogenic ultracentrifugation and characterized by transmission electron microscopy, Western blotting and nanoparticle tracking analysis. HRECs were randomly divided into a normal control group (group A), a high glucose model group (group B), a high glucose group with 25 μg/mL (group C), 50 μg/mL (group D), and 100 μg/mL exosomes (group E). Twenty-four hours after coculture, the cell proliferation rate was detected using flow cytometry, and the VEGF-A level was detected using immunofluorescence. After coculture 8, 16, and 24h, the expression levels of VEGF-A in each group were detected using PCR and Western blots. RESULTS: The characteristic morphology (membrane structured vesicles) and size (diameter between 50 and 200 nm) were observed under transmission electron microscopy. The average diameter of 122.7 nm was discovered by nanoparticle tracking analysis (NTA). The exosomal markers CD9, CD63, and HSP70 were strongly detected. The proliferation rate of the cells in group B increased after 24h of coculture. Immunofluorescence analyses revealed that the upregulation of VEGF-A expression in HRECs stimulated by high glucose could be downregulated by cocultured hUCMSC-derived exosomes (F=39.03, P<0.01). The upregulation of VEGF-A protein (group C: F=7.96; group D: F=17.29; group E: F=11.89; 8h: F=9.45; 16h: F=12.86; 24h: F=42.28, P<0.05) and mRNA (group C: F=4.137; group D: F=13.64; group E: F=22.19; 8h: F=7.253; 16h: F=16.98; 24h: F=22.62, P<0.05) in HRECs stimulated by high glucose was downregulated by cocultured hUCMSC-derived exosomes (P<0.05). CONCLUSION: hUCMSC-derived exosomes downregulate VEGF-A expression in HRECs stimulated by high glucose in time and concentration dependent manner.

scholarly journals In Vitro Construction of Scaffold-Free Bilayered Tissue-Engineered Skin Containing Capillary Networks

2013 ◽  
Vol 2013 ◽  
pp. 1-8 ◽  
Author(s):  
Yuan Liu ◽  
Hailang Luo ◽  
Xinwen Wang ◽  
Akimichi Takemura ◽  
Yi Ru Fang ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Endothelial Cells ◽  
Dermal Fibroblasts ◽  
Microvascular Endothelial Cells ◽  
Skin Substitutes ◽  
Tem Analysis ◽  
Capillary Networks ◽  
Transmission Electron

Many types of skin substitutes have been constructed using exogenous materials. Angiogenesis is an important factor for tissue-engineered skin constructs. In this study, we constructed a scaffold-free bilayered tissue-engineered skin containing a capillary network. First, we cocultured dermal fibroblasts with dermal microvascular endothelial cells at a ratio of 2 : 1. A fibrous sheet was formed by the interactions between the fibroblasts and the endothelial cells, and capillary-like structures were observed after 20 days of coculture. Epithelial cells were then seeded on the fibrous sheet to assemble the bilayered tissue. HE staining showed that tissue-engineered skin exhibited a stratified epidermis after 7 days. Immunostaining showed that the epithelium promoted the formation of capillary-like structures. Transmission electron microscopy (TEM) analysis showed that the capillary-like structures were typical microblood vessels. ELISA demonstrated that vascularization was promoted by significant upregulation of vascularization associated growth factors due to interactions among the 3 types of cells in the bilayer, as compared to cocultures of fibroblast and endothelial cells and monocultures.

scholarly journals Effect of Electrocautery on Endothelial Integrity of the Internal Thoracic Artery: Ultrastructural Analysis with Transmission Electron Microscopy

2014 ◽  
Vol 41 (5) ◽  
pp. 484-490 ◽  
Author(s):  
Burak Onan ◽  
Mehmet Yeniterzi ◽  
Ismihan Selen Onan ◽  
Burak Ersoy ◽  
Suheyla Gonca ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Endothelial Cells ◽  
Internal Thoracic Artery ◽  
Ultrastructural Changes ◽  
Control Group ◽  
Elastic Tissue ◽  
Cytoplasmic Organelles ◽  
Transmission Electron ◽  
Sharp Dissection

The internal thoracic artery (ITA) is typically harvested from the chest wall by means of conventional electrocautery. We investigated the effects of electrocautery on endothelial-cell and vessel-wall morphology at the ultrastructural level during ITA harvesting. Internal thoracic artery specimens from 20 patients who underwent elective coronary artery bypass grafting were investigated in 2 groups. The ITA grafts were sharply dissected with use of a scalpel and clips in the control group (n=10) and were harvested by means of electrocautery in the study group (n=10). Each sample was evaluated for intimal, elastic-tissue, muscular-layer, and adventitial changes. Free flow was measured intraoperatively. Light microscopic examinations were performed after hematoxylin-eosin and Masson's trichrome staining. Transmission electron microscopy was used to evaluate ultrastructural changes in the endothelial cells and vessel walls of each ITA. In the sharp-dissection group, the endothelial surfaces were lined with normal amounts of original endothelium, endothelial cells were distinctly attached to the basal lamina, cytoplasmic organelles were evident, and intercellular junctional complexes were intact. Conversely, in the electrocautery group, the morphologic integrity of endothelial cells was distorted, with some cell separations and splits, contracted cells, numerous large cytoplasmic vacuoles, and no visible cytoplasmic organelles. The subendothelial layer exhibited disintegration. Free ITA flow was higher in the sharp-dissection group (P=0.04). The integrity of endothelial cells can be better preserved when the ITA is mobilized by means of sharp dissection, rather than solely by electrocautery; we recommend a combined approach.

scholarly journals Handling Cell Culture Monolayers for Transmission Electron Microscopy

2013 ◽  
Vol 21 (3) ◽  
pp. 36-39 ◽  
Author(s):  
Leona Cohen-Gould
Keyword(s):  
Electron Microscopy ◽  
Skeletal Muscle ◽  
Cell Culture ◽  
Transmission Electron Microscopy ◽  
Cells In Culture ◽  
Area Of Interest ◽  
Transmission Electron

In our Core Facility, we are presented with a wide variety of tissue and sample types to process for TEM observation. Some tissues, such as liver, do not present orientation problems. Other tissues such as skeletal muscle have specific orientations that must be maintained. As long as the initial dissection of the tissue is done so that the orientation or area of interest can be identified, these samples do not present a problem. Likewise, when it is not necessary to maintain orientation, cells in culture can be treated with trypsin, to release them from the plate, and pelleted. Or the monolayer can be fixed in vitro and then scraped and pelleted, and the pellet can be processed while adherent to the wall of the Eppendorf tube or handled like a small piece of tissue, among other methods.

Pharmaceutical Research Applications of Transmission Electron Microscopy of Mammalian Cells in Culture

2008 ◽  
Vol 14 (S2) ◽  
pp. 164-165
Author(s):  
JR Megill
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
New Mexico ◽  
Mammalian Cells ◽  
Pharmaceutical Research ◽  
Cells In Culture ◽  
Transmission Electron

Extended abstract of a paper presented at Microscopy and Microanalysis 2008 in Albuquerque, New Mexico, USA, August 3 – August 7, 2008

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