scholarly journals Potential Effects of Nonadherent on Adherent Human Umbilical Venous Endothelial Cells in Cell Culture

2021 ◽  
Vol 22 (3) ◽  
pp. 1493
Author(s):  
Christian Schulz ◽  
Anne Krüger-Genge ◽  
Andreas Lendlein ◽  
Jan-Heiner Küpper ◽  
Friedrich Jung
Keyword(s):  
Endothelial Cells ◽  
Culture Medium ◽  
Membrane Integrity ◽  
Time Dependent ◽  
Implant Materials ◽  
And Function ◽  
Nonadherent Cells ◽  
Initial Cell Adhesion ◽  
Dependent Behaviour

The adherence and shear-resistance of human umbilical venous endothelial cells (HUVEC) on polymers is determined in vitro in order to qualify cardiovascular implant materials. In these tests, variable fractions of HUVEC do not adhere to the material but remain suspended in the culture medium. Nonadherent HUVEC usually stop growing, rapidly lose their viability and can release mediators able to influence the growth and function of the adherent HUVEC. The aim of this study was the investigation of the time dependent behaviour of HUVEC under controlled nonadherent conditions, in order to gain insights into potential influences of these cells on their surrounding environment in particular adherent HUVEC in the context of in vitro biofunctionality assessment of cardiovascular implant materials. Data from adherent or nonadherent HUVEC growing on polystyrene-based cell adhesive tissue culture plates (TCP) or nonadhesive low attachment plates (LAP) allow to calculate the number of mediators released into the culture medium either from adherent or nonadherent cells. Thus, the source of the inflammatory mediators can be identified. For nonadherent HUVEC, a time-dependent aggregation without further proliferation was observed. The rate of apoptotic/dead HUVEC progressively increased over 90% within two days. Concomitant with distinct blebbing and loss of membrane integrity over time, augmented releases of prostacyclin (PGI2, up to 2.91 ± 0.62 fg/cell) and platelet-derived growth factor BB (PDGF-BB, up to 1.46 ± 0.42 fg/cell) were detected. The study revealed that nonadherent, dying HUVEC released mediators, which can influence the surrounding microenvironment and thereby the results of in vitro biofunctionality assessment of cardiovascular implant materials. Neglecting nonadherent HUVEC bears the risk for under- or overestimation of the materials endothelialization potential, which could lead to the loss of relevant candidates or to uncertainty with regard to their suitability for cardiac applications. One approach to minimize the influence from nonadherent endothelial cells could be their removal shortly after observing initial cell adhesion. However, this would require an individual adaptation of the study design, depending on the properties of the biomaterial used.

scholarly journals Venous and Arterial Endothelial Cells from Human Umbilical Cords: Potential Cell Sources for Cardiovascular Research

2021 ◽  
Vol 22 (2) ◽  
pp. 978
Author(s):  
Skadi Lau ◽  
Manfred Gossen ◽  
Andreas Lendlein ◽  
Friedrich Jung
Keyword(s):  
Endothelial Cells ◽  
Umbilical Cord ◽  
Membrane Integrity ◽  
Cell Types ◽  
Cell Number ◽  
Human Umbilical Cord ◽  
Cardiovascular Research ◽  
Cardiovascular Devices ◽  
Arterial Endothelial Cells

Although cardiovascular devices are mostly implanted in arteries or to replace arteries, in vitro studies on implant endothelialization are commonly performed with human umbilical cord-derived venous endothelial cells (HUVEC). In light of considerable differences, both morphologically and functionally, between arterial and venous endothelial cells, we here compare HUVEC and human umbilical cord-derived arterial endothelial cells (HUAEC) regarding their equivalence as an endothelial cell in vitro model for cardiovascular research. No differences were found in either for the tested parameters. The metabolic activity and lactate dehydrogenase, an indicator for the membrane integrity, slightly decreased over seven days of cultivation upon normalization to the cell number. The amount of secreted nitrite and nitrate, as well as prostacyclin per cell, also decreased slightly over time. Thromboxane B2 was secreted in constant amounts per cell at all time points. The Von Willebrand factor remained mainly intracellularly up to seven days of cultivation. In contrast, collagen and laminin were secreted into the extracellular space with increasing cell density. Based on these results one might argue that both cell types are equally suited for cardiovascular research. However, future studies should investigate further cell functionalities, and whether arterial endothelial cells from implantation-relevant areas, such as coronary arteries in the heart, are superior to umbilical cord-derived endothelial cells.

Isolation, Partial Purification and Characterisation of A Plasminogen Activator Produced by Endothelial Cells in Vitro

1979 ◽  
Author(s):  
W.E. Laug
Keyword(s):  
Endothelial Cells ◽  
Proteolytic Activity ◽  
Culture Medium ◽  
Partial Purification ◽  
Diisopropyl Fluorophosphate ◽  
Cell Lysate ◽  
Endothelial Cell Cultures ◽  
Ph Range ◽  
Confluent Endothelial Cell

Cloned endothelial cells obtained from the aorta of 1-2 day old calves produced high fibrinolytic activity, which was 90% dependent upon the presence of plasminogen when grown on 125 I fibrin coated dishes. High plasminogen-dependent proteolytic activity was also demonstrated in the cell lysate and in the culture medium of the cells. The production and secretion of this prtitease were found to increase during the log phase of cell growth and to reach a maximum at con fluency. Thereafter they remained constantly high. This protease, partially purified from the culture medium of confluent endothelial cell cultures, is aiginine specific and activates plasminogen by piOteolytic cleavage to plasmin. Its proteolytic activity which is highest in the pH range of 7.5 to 8.0 is irreversibly inhibited by diisopropyl fluorophosphate, suggesting that it is a serine protease. The molecular weight of this protease is approximately S2000.

scholarly journals Plasminogen activator inhibitor-1 secretion of endothelial cells increases fibrinolytic resistance of an in vitro fibrin clot: evidence for a key role of endothelial cells in thrombolytic resistance

Blood ◽  
1996 ◽  
Vol 87 (10) ◽  
pp. 4204-4213 ◽  
Author(s):  
S Handt ◽  
WG Jerome ◽  
L Tietze ◽  
RR Hantgan
Keyword(s):  
Endothelial Cells ◽  
Blood Vessels ◽  
Plasminogen Activator ◽  
Plasminogen Activator Inhibitor ◽  
Time Dependent ◽  
Plasminogen Activator Inhibitor 1 ◽  
Necrosis Factor Alpha ◽  
Activator Inhibitor ◽  
Pai 1

Time-dependent thrombolytic resistance is a critical problem in thrombolytic therapy for acute myocardial infarction. Platelets have been regarded as the main source of plasminogen activator inhibitor-1 (PAI-1) found in occlusive platelet-rich clots. However, endothelial cells are also known to influence the fibrinolytic capacity of blood vessels, but their ability to actively mediate time-dependent thrombolytic resistance has not been fully established. We will show that, in vitro, tumor necrosis factor-alpha-stimulated endothelial cells secrete large amounts of PAI-1 over a period of hours, which then binds to fibrin and protects the clot from tissue plasminogen activator- induced fibrinolysis. In vivo, endothelial cells covering atherosclerotic plaques are influenced by cytokines synthesized by plaque cells. Therefore, we propose that continuous activation of endothelial cells in atherosclerotic blood vessels, followed by elevated PAI-1 secretion and storage of active PAI-1 in the fibrin matrix, leads to clot stabilization. This scenario makes endothelial cells a major factor in time-dependent thrombolytic resistance.

Does prostacyclin (PGI2) support porcine corpus luteum function? – data from in vitro study

2016 ◽  
Vol 62 (5) ◽  
pp. 49
Author(s):  
Magdalena Julia Szymańska ◽  
Agnieszka Blitek
Keyword(s):  
Endothelial Cells ◽  
Mrna Expression ◽  
Corpus Luteum ◽  
Estrous Cycle ◽  
In Vitro Study ◽  
Luteal Cells ◽  
Enzymatic Isolation ◽  
Effective Doses ◽  
And Function

Background. Prostacyclin (PGI2) of luteal origin is involved in the control of corpus luteum (CL) development and function in cattle. PGI2 may regulate the process of angiogenesis and may stimulate progesterone (P4) secretion by luteal cells via its specific receptors, PTGIR. In contrast to cattle, the role of PGI2 in the pig CL has not yet been described.Aim. The present study aimed to investigate the effect of PGI2 on 1) P4 secretion by luteal cells, and 2) the expression of angiogenesis-related genes in endothelial cells of the porcine CL.Methods. CL collected from gilts on day 5-7 of the estrous cycle were used for enzymatic isolation of luteal (Experiment 1) and endothelial (Experiment 2) cells. In Exp. 1, cultured luteal cells were incubated with increasing (0, 0.01, 0.1, 1, 5 µM) doses of PGI2 analogues: iloprost (ILO) and carbaprostacyclin (cPGI2) for 8 h. To determine the effective doses of PGI2 analogues, P4 concentration in culture medium was examined by RIA. Thereafter, luteal cells were treated with ILO and cPGI2 at the concentration of 1 and 5 µM in the presence or absence of PTGIR antagonist (CAY10441). After 8 h of incubation the medium was collected for P4 determination. In Exp. 2, isolated endothelial cells were treated for 24 h with ILO and cPGI2 at doses of 1 and 5 µM. Then, cells were collected for analysis of Ang-1 and -2 mRNA expression using qPCR.Results. Both, ILO and cPGI2 affected P4 secretion by luteal cells. Elevated levels of P4 were observed in medium after treatment of luteal cells with 1 µM of ILO and 0.1, 1 and 5 µM of cPGI2 compared with control values (p<0.05). The addition of CAY10441 inhibited the stimulatory effect of ILO on P4 secretion, while did not change P4 production by luteal cells incubated with cPGI2. Moreover, PGI2 analogues differentially affected (p<0.05) the expression of proangiogenic factors. ILO stimulated Ang-2, whereas cPGI2 positively affected Ang-1 mRNA expression in endothelial cells at concentrations of 1 µM and 5 µM, respectively.Conclusion. PGI2 affects P4 secretion during luteal phase of the estrous cycle and may regulate the process of angiogenesis in the porcine CL.

Wall Shear Stress Measurements in an Arterial Flow Bioreactor

2011 ◽  
Author(s):  
Elizabeth Voigt ◽  
Cara Buchanan ◽  
Jaime Schmieg ◽  
M. Nichole Rylander ◽  
Pavlos Vlachos
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Vascular System ◽  
Vascular Endothelial Cells ◽  
Shear Stresses ◽  
Bulk Flow Rate ◽  
Wall Shear ◽  
Endothelial Cell Response ◽  
And Function

Physiological flow parameters such as pressure and stress inside the vascular system strongly influence the physiology and function of vascular endothelial cells [1]. Variations in the shear stress experienced by endothelial cells affect morphology, alignment with the flow, mechanical strength, rate of proliferation, and gene expression [2]. Although it is known that these factors are dependent on the hemodynamics of the flow, the relationship has not been accurately quantified. In vitro bioreactor flow loops have been developed to simulate vascular flow for tissue conditioning and measurement of the endothelial cell response to varying shear [3–5]; however, wall shear stresses (WSS) have been estimated from the bulk flow rate by assuming Poiseuille flow [2, 6]. Due to the pulsatility of the flow, biochemical interactions, and the typically short vessel length, this assumption is fundamentally incorrect; however, the level of inaccuracy has not been quantified.

ENGINEERING OF SURFACE FUNCTIONALITY ONTO POLYSTYRENE MICROCARRIERS FOR THE ATTACHMENT AND GROWTH OF HUMAN ENDOTHELIAL CELLS

2014 ◽  
Vol 02 (03n04) ◽  
pp. 1450003 ◽  
Author(s):  
GORDON M. XIONG ◽  
JOHN S. FOORD ◽  
JON-PAUL GRIFFITHS ◽  
EMILY M. PARKER ◽  
MARK G. MOLONEY ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Cellular Adhesion ◽  
Human Endothelial Cells ◽  
Surface Functionality ◽  
Microcarrier Beads ◽  
Umbilical Vein Endothelial Cells ◽  
And Function ◽  
Cell Carriers ◽  
Initial Cell Adhesion

This work reports the effects of introducing diverse chemical functionalities onto the surface of polystyrene microcarrier beads on their ability to function as injectable cell carriers. Cellular adhesion and proliferation, as well as cellular outgrowths from microcarrier surfaces, using human umbilical vein endothelial cells (HUVECs), were examined in detail. It was observed that initial cell adhesion appeared to be most significantly decreased by hydrophobicity, whilst cell proliferation appeared to be improved in most chemical functional groups over unmodified polystyrene. Overall, our study highlights the importance of surface chemistry in directing the growth and function of human endothelial cells.

Defective Peripheral Blood Endothelial Cell Progenitors in Patients with Scleroderma and Psoriatic Arthritis

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 4724-4724
Author(s):  
Taxiarchis V Kourelis ◽  
Akrivi D Manola ◽  
Despoina Adamidou ◽  
Lazaros Sakkas ◽  
Eyagelia Mperou ◽  
...  
Keyword(s):  
Psoriatic Arthritis ◽  
Cord Blood ◽  
Peripheral Blood ◽  
Culture Medium ◽  
Autologous Bone Marrow ◽  
Autologous Serum ◽  
Cd34 Cells ◽  
Cord Blood Cd34 ◽  
And Function

Abstract Vasculogenesis is known to be defective in patients with scleroderma (SS) and psoriatic arthritis (PA) with vessel loss in the former and hypertrophic blood vessels in the latter in affected areas. We studied the number and function of peripheral blood endothelial progenitors (PBEP) in patients with SS and PA to elucidate the mechanism of EC dysfunction. Materials and Methods: Eleven patients with SS, 13 patients with PA and 7 healthy individuals were studied. We measured CD133+/CD146+ cells in peripheral blood (PB) by immunofluorescence. We performed cell cultures of isolated CD34+ cells in endocult medium and examined the endothelial colonies (EPC). We also performed cocultures of CD34+ and autologous bone marrow stromal cells (BMSC) in double chambers. We also performed cocultures of BMSC from patients and normal endothelial cells from cord blood. Results: The number of CD133+/CD146+ cell in PB was increased the 2 groups of patients compared to controls. The number of EC colonies in endocult did not differ in the 3 groups. The presence of autologous serum within the culture medium reduced the number of colonies in 3 patients with SS. The number and the size of EC colonies from SS patients in vitro were significantly reduced (p&lt;0.01) after co-cultures of autologous BMSC with CD34+ in culture plates with insert while they were increased (p&lt;0.01) from patients with PA. The same was true when cord blood CD34+ cells were cultured in endocult medium in the presence of BMSC of SS and PA patients. Conclusion: EC progenitors from patients with SS and PA are increased in PB and develop normal EC colonies in vitro. They developed decreased colonies in SS and increased colonies in PA when cultured together with with autologous BMSC. This means that possible cell-cell or humoral interactions between EC and some cellular component within BMSC affect the survival and differentiation of EC.

Adenosine and hypoxia stimulate proliferation and migration of endothelial cells

1988 ◽  
Vol 255 (3) ◽  
pp. H554-H562 ◽  
Author(s):  
C. J. Meininger ◽  
M. E. Schelling ◽  
H. J. Granger
Keyword(s):  
Cell Proliferation ◽  
Endothelial Cells ◽  
Culture Medium ◽  
Culture Conditions ◽  
Hypoxic Conditions ◽  
Cell Counts ◽  
And Migration ◽  
Stimulate Cell Proliferation ◽  
Proliferation And Migration

The proliferation of bovine aortic or coronary venular endothelial cells (EC) in vitro was stimulated by the addition of adenosine (0.5 or 5.0 microM) to the culture medium. Cell counts of adenosine-treated aortic EC were 23–76% and coronary venular EC 19–52% greater than nontreated controls. Because adenosine is known to be released by hypoxic tissues, cell proliferation was quantitated when aortic EC were grown at 2% O2. Cell counts were 41–102% greater under hypoxic conditions than when cells were grown at standard tissue culture conditions (approximately 20% O2). When culture medium conditioned by coronary EC grown at 2% O2 was added to EC growing at standard conditions, cell counts were 24–69% greater than controls with medium conditioned by coronary EC grown at 20% O2. This suggests that hypoxia causes endothelial cells to release a factor(s) into the medium that can stimulate cell proliferation. The addition of the adenosine receptor blocker 8-phenyltheophylline (10(-5) M) prevented the stimulation of proliferation caused by hypoxia-conditioned medium, 2% O2 or 5.0 microM adenosine, suggesting that adenosine mediates its effect via an external membrane receptor. Adenosine also stimulated EC chemotaxis. Taken together, these results suggest that adenosine, released as a result of tissue hypoxia, may act as an angiogenic stimulus for the growth of new blood vessels.

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