Endothelial Seeding of Vascular Prostheses: A Technique of In Situ Enzymatic Retrieval of Endothelial Cells Without Vein Sacrifice

SummaryPurpose: Successful development of a vascular prosthesis lined with endothelial cells (EC) may depend on the ability of the attached cells to resist shear forces after implantation. The present study was designed to investigate EC detachment from extracellular matrix (ECM) precoated vascular prostheses, caused by shear stress in vitro and to test the performance of these grafts in vivo. Methods: Bovine aortic endothelial cells were seeded inside untreated polytetrafluoro-ethylene (PTFE) vascular graft (10 X 0.6 cm), PTFE graft precoated with fibronectin (FN), or PTFE precoated with FN and a naturally produced ECM (106 cells/graft). Sixteen hours after seeding the medium was replaced and unattached cells counted. The strength of endothelial cell attachment was evaluated by subjecting the grafts to a physiologic shear stress of 15 dynes/cm2 for 1 h. The detached cells were collected and quantitated. PTFE or EC preseeded ECM coated grafts were implanted in the common carotid arteries of dogs. Results: While little or no differences were found in the extent of endothelial cell attachment to the various grafts (79%, 87% and 94% of the cells attached to PTFE, FN precoated PTFE, or FN+ECM precoated PTFE, respectively), the number of cells retained after a shear stress was significanly increased on ECM coated PTFE (20%, 54% and 85% on PTFE, FN coated PTFE, and FN+ECM coated PTFE, respectively, p <0.01). Implantation experiments in dogs revealed a significant increase in EC coverage and a reduced incidence of thrombus formation on ECM coated grafts that were seeded with autologous saphenous vein endothelial cells prior to implantation. Conclusion: ECM coating significantly increased the strength of endothelial cell attachment to vascular prostheses subjected to shear stress. The presence of adhesive macromolecules and potent endothelial cell growth promoting factors may render the ECM a promising substrate for vascular prostheses.

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Mechanism of acetylcholine-induced Ca2+ oscillation with high frequency in individual endothelial cells in rat aorta in situ

The Japanese Journal of Pharmacology ◽

10.1016/s0021-5198(19)40707-5 ◽

1998 ◽

Vol 76 ◽

pp. 149

Author(s):

Gousei Lee ◽

Hisayuki Qhata ◽

Yosuke Ujike ◽

Chieko Yanagi ◽

Kazutaka Momose

Keyword(s):

Endothelial Cells ◽

High Frequency ◽

Rat Aorta

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Selective biological response of human pulmonary microvascular endothelial cells and human pulmonary artery smooth muscle cells on cold-plasma-modified polyester vascular prostheses

Biomedical Materials ◽

10.1088/1748-6041/6/6/065003 ◽

2011 ◽

Vol 6 (6) ◽

pp. 065003 ◽

Cited By ~ 8

Author(s):

N Blanchemain ◽

M R Aguilar ◽

F Chai ◽

M Jimenez ◽

E Jean-Baptiste ◽

...

Keyword(s):

Endothelial Cells ◽

Smooth Muscle ◽

Pulmonary Artery ◽

Smooth Muscle Cells ◽

Cold Plasma ◽

Biological Response ◽

Microvascular Endothelial Cells ◽

Vascular Prostheses ◽

Pulmonary Artery Smooth Muscle ◽

Human Pulmonary Artery

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43. Determination of cryoprotectant permeability properties in monolayers of bovine endothelial cells using an in situ fluorescence quenching technique

Cryobiology ◽

10.1016/j.cryobiol.2009.10.057 ◽

2009 ◽

Vol 59 (3) ◽

pp. 382

Author(s):

A.K. Fry ◽

A.Z. Higgins

Keyword(s):

Endothelial Cells ◽

Fluorescence Quenching ◽

Bovine Endothelial Cells ◽

Fluorescence Quenching Technique

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The quest for endothelial atypical cannabinoid receptor: BKCa channels act as cellular sensors for cannabinoids in in vitro and in situ endothelial cells

Vascular Pharmacology ◽

10.1016/j.vph.2018.01.004 ◽

2018 ◽

Vol 102 ◽

pp. 44-55 ◽

Cited By ~ 8

Author(s):

Alexander I. Bondarenko ◽

Olga Panasiuk ◽

Konstantin Drachuk ◽

Fabrizio Montecucco ◽

Karim J. Brandt ◽

...

Keyword(s):

Endothelial Cells ◽

Cannabinoid Receptor ◽

Bkca Channels ◽

Cellular Sensors

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The relationship between cell proliferation and the transcription of the nuclear oncogenes c-myc, c-myb and c-ets-1 during feather morphogenesis in the chick embryo

Development ◽

10.1242/dev.111.3.699 ◽

1991 ◽

Vol 111 (3) ◽

pp. 699-713 ◽

Cited By ~ 1

Author(s):

X. Desbiens ◽

C. Queva ◽

T. Jaffredo ◽

D. Stehelin ◽

B. Vandenbunder

Keyword(s):

Cell Proliferation ◽

Endothelial Cells ◽

Spatial Distribution ◽

Chick Embryo ◽

Expression Patterns ◽

S Phase ◽

Skin Morphogenesis ◽

Dermal Cells ◽

The Relationship

We have described the expression of three nuclear protooncogenes, c-myc, c-myb and c-ets-1 during feather morphogenesis in the chick embryo. In parallel with the expression patterns obtained by in situ hybridization, we have mapped the spatial distribution of S-phase cells by monitoring the incorporation of 5-bromodeoxyuridine. We do not detect c-myc or c-myb transcripts during the early stages when S-phase cells are scattered in the dermis and in the epidermis. Rather c-ets-1 transcripts are abundant in the dermal cells which divide and accumulate under the uniform epidermis. At the onset of the formation of the feather bud, cells within each rudiment cease DNA replicative activities and c-myc transcripts are detected both in the epidermis and in the underlying dermis. This expression precedes the reentry into the S phase. The transcription of c-myb, which has been previously tightly linked to hemopoietic cells is also detected in the developing skin. This expression is essentially located in proliferating epidermal cells on and after the beginning of feather outgrowth. As feather outgrowth proceeds, the distribution of c-myc and c-myb transcripts is restricted to the highly proliferating epidermis. In contrast c-ets-1 transcripts are never detected in the epidermis. During the later stages of skin morphogenesis, the transcription of c-ets-1 is restricted to the endothelial cells of blood vessels, as previously described. We suggest that the differential expression of these nuclear oncogenes reflects the activation of different mitotic controlling pathways during the development of the skin.

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Endothelial Cell Seeding on PTFE Vascular Prostheses Using a Standardized Seeding Technique

The International Journal of Artificial Organs ◽

10.1177/039139888901200410 ◽

1989 ◽

Vol 12 (4) ◽

pp. 270-275 ◽

Cited By ~ 3

Author(s):

J. Gerlach ◽

K.-M. Kreusel ◽

H.H. Schauwecker ◽

E.S. Bücherl

Keyword(s):

Endothelial Cells ◽

Rotation Speed ◽

Vascular Grafts ◽

Cell Number ◽

Cell Seeding ◽

Vascular Prostheses ◽

Adhesion Rate ◽

Endothelial Cell Seeding ◽

Kinetics Of ◽

Graft Diameter

A standardized method was developed for seeding endothelial cells (EC) in tubular vascular grafts. A rotational cell seeding device for tubular prostheses is presented and parameters influencing the kinetics of cell adhesion (rotation speed, graft diameter, cell suspension level, inoculated cell number) are reported. Seeding EC in 14 mm ID PTFE vascular grafts with rotation rate of 10 rph gave an adhesion rate of 80% in a homogeneous monolayer.

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Tenascin in rat lung development: in situ localization and cellular sources

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1995.269.4.l482 ◽

1995 ◽

Vol 269 (4) ◽

pp. L482-L491 ◽

Cited By ~ 7

Author(s):

Y. Zhao ◽

S. L. Young

Keyword(s):

Extracellular Matrix ◽

Endothelial Cells ◽

Epithelial Cells ◽

Lung Tissue ◽

Lung Development ◽

Alveolar Epithelial Cells ◽

Lung Fibroblasts ◽

Rat Lung ◽

Alveolar Epithelial

Tenascin (TN) is a hexameric extracellular matrix glycoprotein that may play an important role during lung development. TN protein is temporally and spatially restricted during lung organogenesis. The temporo-spatial and cellular expression of TN mRNA in lung remains unclear. Localization of message expression of TN in rat lung tissue was first investigated by using in situ hybridization performed with an antisense RNA probe. TN mRNA was present primarily within the mesenchyme of day 16 gestational age fetal rat lung tissue, whereas immunoreactive TN protein was found along the basement membrane. In postnatal day 3 rat lung tissue, TN mRNA was detected along alveolar septal walls and was concentrated at secondary septal tips. Expression of TN message was consistent with localization of immunoreactive TN protein. Accumulation of TN mRNA in alveolar septal tips suggests that mesenchyme may be the major source of TN mRNA. To investigate the cellular source of TN in rat lung, we studied the expression of TN in cultured rat lung fibroblasts, endothelial cells, and alveolar epithelial cells. Two TN isoforms having molecular mass of 230 and 180 kDa were in conditioned medium and in cellular extracts of lung fibroblasts and endothelial cells. TN was secreted and deposited in the extracellular matrix closely associated with the surface of lung fibroblasts and endothelial cells. Lung alveolar epithelial cells showed undetectable or barely detectable amounts of TN. These studies demonstrated that TN isoforms are expressed not only by lung fibroblasts but also by lung endothelial cells. The unique spatial localization of TN mRNA during lung development and expression of TN by different lung cell types suggested TN may be involved in matrix organization and cell-cell interactions during lung development.

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Limitations of In-Vitro Labeling of Endothelial Cells with Indium-111 Oxine

Cell Transplantation ◽

10.1177/096368979500400307 ◽

1995 ◽

Vol 4 (3) ◽

pp. 291-296 ◽

Cited By ~ 6

Author(s):

H.M.H. Carr ◽

J.V. Smyth ◽

O.B. Rooney ◽

P.D. Dodd ◽

H. Sharma ◽

...

Keyword(s):

Endothelial Cells ◽

Endothelial Cell ◽

Cell Attachment ◽

Control Group ◽

Vascular Prostheses ◽

Long Term Effect ◽

Indium 111 ◽

Suitable Technique

Indium-111 oxine labeling is widely used as a marker of endothelial cell attachment to vascular prostheses. The long term effect of labeling human adult endothelial cells (HAECs) with this isotope has not been determined. In this study the viability of labeled HAECs, leakage of isotope from labeled cells and adherence of circulating isotope to fibronectin coated prostheses were investigated over 24 h. The effect of incubation time on labeling efficiency was also assessed. There were significant differences in cell viability between the labeled and unlabeled groups beyond 4 h (p < 0.005, 2-tailed, unpaired t-test). In the control group cell numbers increased by 42% while in the labeled group this had decreased by 20% at 24 h. Spontaneous leakage increased with time but was maximal in the first 2 h. Adherence of circulating isotope to fibronectin coated expanded polytetrafluoroethylene (ePTFE) grafts was minimal but was significantly greater to gelatin impregnated Dacron (GEL-SEAL) beyond 1 hour (p < 0.05). Incubation times greater than 5 minutes during labeling do not significantly improve labeling efficiency, and may contribute to toxicity by prolonging exposure to oxine. Indium-111 oxine labeling of HAECs is a suitable technique for acute studies of endothelial cell kinetics up to 4 h, but its use in chronic studies may lead to significant underestimations of cell retention.

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