Harvest Of Endothelial Cells Research Articles

Abstract 13577: Burden of Cardiometabolic Risk Factors and Vascular Health

Introduction: Cardiometabolic risk factors (diabetes, obesity, and hypertension) are highly prevalent and contribute to cardiovascular disease (CVD). Endothelial dysfunction often precedes CVD development. Direct brachial vein endothelial cell (EC) harvesting enables the examination of vascular health via assessment of EC gene expression. Hypothesis: We hypothesized that individuals with greater cardiometabolic risk would demonstrate a more pro-inflammatory EC transcriptome. Methods: Adult participants without CVD underwent EC harvesting followed by RNA sequencing. We categorized individuals by number of risk factors (hypertension, diabetes, obesity) as 0 through 3. To evaluate the association between risk factor burden and outcome transcripts, we performed multivariable linear regression, adjusting for age, sex, and race/ethnicity. Results: Among 18 individuals (mean age 47 ± 14, 44% female, and 61% white), 17%, 44%, 28%, and 11% had 0,1,2, and 3 prevalent risk factors, respectively. EC RNA sequencing revealed 588 differentially expressed transcripts (p-adj <0.05) with 585 upregulated, 3 downregulated, and excellent clustering ( Fig. a, b ). Gene ontology enrichment analysis demonstrated enriched and upregulated pathways of T-cell activation (NES = 2.22, p<0.001), leukocyte differentiation (NES= 2.16, p<0.001), and platelet aggregation (NES= 1.66, p=0.03) and downregulated pathways of endothelial cell proliferation (NES= -1.68, p=0.03). ( Fig. c ) Select upregulated genes (log 2FC >3, p-adj<0.05) included VCAM1, CEACAM1, ADAM 17, and CD99L2, with a graded increase in mean normalized counts with increasing number of risk factors. ( Fig. d ) Conclusions: We demonstrate a proinflammatory, pro-adhesive, and pro-coagulant EC transcriptome associated with increased cardiometabolic risk factor burden offering insight into potential mechanisms linking cardiometabolic risk factors with the development of CVD.

Abstract 227: Statins Decrease Endothelial Cell Inflammation On Top Of Maximal Cholesterol Reduction: Findings From The American Heart Association Strategically Focused Cardiometabolic Health And Type 2 Diabetes Mellitus Research Network

Background: Dyslipidemia induces a proinflammatory endothelium and vascular dysfunction. Lipid lowering therapy (LLT) consistently reduces cardiovascular (CV) events. Clinical studies suggest that statin-based LLT has pleiotropic anti-inflammatory effects beyond cholesterol reduction. However, the impact of statins or ezetimibe on top of potent cholesterol reduction is unknown. Thus, on a background of PCSK9 inhibition (with evolocumab), we analyzed the effect of LLT with atorvastatin or ezetimibe on the endothelium using a novel direct brachial vein (BV) endothelial cell (EC) harvesting technique. Methods: CHORD (CHOlesterol lowering and Residual Risk in Diabetes) is an ongoing, prospective study designed to evaluate the effect of maximal LLT on CV risk. Subjects with LDL-C >100mg/dl were treated with evolocumab (140mg/2 weeks) and either atorvastatin (80mg/day) or ezetimibe (10mg/day) for 30 days. In a subset of participants, EC harvesting was performed by inserting a J-wire into the BV, and ECs were isolated and analyzed using next generation RNA sequencing. Results: Among 20 participants (11 atorvastatin, 9 ezetimibe) undergoing EC RNASeq, median LDL-C (130 mg/dl, IQR [110, 147]) decreased by 75% [67, 81] on background PCSK9 inhibition with no significant difference in 30-day LDL-C between atorvastatin (27 mg/dl [20, 32]) and ezetimibe (41 mg/dl [26, 53]) groups (p=0.07). Following cholesterol reduction, 850 genes were upregulated and 2468 were downregulated (p<0.05) with downregulation of eNOS, Endothelin-1, and Senescence pathways by canonical pathway analysis. When stratified by LLT, patients receiving atorvastatin had a significant decrease in inflammation and senescence pathways compared to patients receiving ezetimibe (Figure). Conclusions: Robust lipid lowering with PCSK9 inhibition plus statins, as opposed to ezetimibe has anti-inflammatory effects and suggests a preferred strategy to improve the endothelium and reduce CV risk.

Harvest of Endothelial Cells from the Balloon Tips of Swan-Ganz Catheters after Right Heart Catheterization.

A variety of pathologies lead to pulmonary hypertension (PH), which is defined as a mean pulmonary artery pressure exceeding 25 mmHg at rest. To further diagnose and manage PH, patients undergo repeated right heart catheterizations (RHC) wherein a Swan-Ganz catheter is advanced into a branch of the pulmonary artery and a balloon is inflated to wedge the catheter tip. This article illustrates a protocol whereby pulmonary artery endothelial cells (PAECs) may be harvested from the balloon tips of Swan-Ganz catheters after RHC, and purified with an anti- CD146 affinity column technique to purify putative PAECs. These cells might be used to provide an in situ snapshot of the biological state of the pulmonary vasculature endothelium to complement hemodynamic measurements obtained during RHC. Harvested and purified PAECs may be used for either cell culture or for subsequent analytical assays such as flow cytometery.

Harvest of Endothelial Cells from the Balloon Tips of Swan-Ganz Catheters after Right Heart Catheterization

Harvest of Endothelial Cells from the Balloon Tips of Swan-Ganz Catheters after Right Heart Catheterization

A novel minimally-invasive method to sample human endothelial cells for molecular profiling.

ObjectiveThe endothelium is a key mediator of vascular homeostasis and cardiovascular health. Molecular research on the human endothelium may provide insight into the mechanisms underlying cardiovascular disease. Prior methodology used to isolate human endothelial cells has suffered from poor yields and contamination with other cell types. We thus sought to develop a minimally invasive technique to obtain endothelial cells derived from human subjects with higher yields and purity.MethodsNine healthy volunteers underwent endothelial cell harvesting from antecubital veins using guidewires. Fluorescence-activated cell sorting (FACS) was subsequently used to purify endothelial cells from contaminating cells using endothelial surface markers (CD34 / CD105 / CD146) with the concomitant absence of leukocyte and platelet specific markers (CD11b / CD45). Endothelial lineage in the purified cell population was confirmed by expression of endothelial specific genes and microRNA using quantitative polymerase chain reaction (PCR).ResultsA median of 4,212 (IQR: 2161 – 6583) endothelial cells were isolated from each subject. Quantitative PCR demonstrated higher expression of von Willebrand Factor (vWF, P<0.001), nitric oxide synthase 3 (NOS3, P<0.001) and vascular cell adhesion molecule 1 (VCAM-1, P<0.003) in the endothelial population compared to similarly isolated leukocytes. Similarly, the level of endothelial specific microRNA-126 was higher in the purified endothelial cells (P<0.001).ConclusionThis state-of-the-art technique isolates human endothelial cells for molecular analysis in higher purity and greater numbers than previously possible. This approach will expedite research on the molecular mechanisms of human cardiovascular disease, elucidating its pathophysiology and potential therapeutic targets.

Endovascular biopsy: Technical feasibility of novel endothelial cell harvesting devices assessed in a rabbit aneurysm model.

The lack of safe and reliable methods to sample vascular tissue in situ limits discovery of the underlying genetic and pathophysiological mechanisms of many vascular disorders, including aneurysms. We investigated the feasibility and comparable efficacy of invivo vascular endothelial cell sampling using a spectrum of endovascular devices. Using the rabbit elastase carotid aneurysm model we evaluated the performance of existing aneurysmal coils, intracranial stents, and stent-like devices to collect vascular endothelial cells. Additionally, we modified a subset of devices to assess the effects of alterations to coil pitch, coil wire contour, and stent surface finishing. Device performance was evaluated by (1) the number of viable endothelial cells harvested, (2) the degree of vascular wall damage analyzed using digital subtraction angiography and histopathological analysis, and (3) the ease of device navigability and retrieval. Isolated cells underwent immunohistochemical analysis to confirm cell type and viability. Coil and stent specifications, technique, and endothelial cell counts were tabulated and statistical analysis performed. Using conventional detachable-type and modified aneurysm coils 11 of 14 (78.6%) harvested endothelial cells with a mean of 7.93 (±8.33) cells/coil, while 15 of 15 (100%) conventional stents, stent-like devices and modified stents harvested endothelial cells with a mean of 831.33 (±887.73) cells/device. Coil stiffness was significantly associated with endothelial cell count in univariate analysis (p = 0.044). For stents and stent-like devices univariate analysis demonstrated stent-to-aorta diameter ratios (p = 0.001), stent length (p = 0.049), and the use of a pulling retrieval technique (p = 0.019) significantly predictive of endothelial cell counts, though a multivariate model using these variables demonstrated only the stent-to-aorta diameter ratio (p = 0.029) predictive of endothelial cell counts. Modified devices did not significantly impact harvesting. The efficacy and safety of existing aneurysm coils, intracranial stents and stent-like devices in collecting viable endothelial cells was confirmed. The technique is reproducible and the quantity and quality of collected endothelial cells is adequate for targeted genetic analysis.

Endovascular biopsy: Technical feasibility of novel endothelial cell harvesting devices assessed in a rabbit aneurysm model

Endovascular biopsy: Technical feasibility of novel endothelial cell harvesting devices assessed in a rabbit aneurysm model

Endovascular Biopsy: Technical Feasibility of Novel Endothelial Cell Harvesting Devices Assessed in a Rabbit Aneurysm Model

Endovascular Biopsy: Technical Feasibility of Novel Endothelial Cell Harvesting Devices Assessed in a Rabbit Aneurysm Model

Prevention of rejection of allogeneic endothelial cells in a biohybrid lung by silencing HLA-class I expression

Variability in Human Leukocyte Antigens (HLA) remains a hurdle to the application of allogeneic cellular products. Due to insufficient autologous endothelial cell harvesting for the biohybrid lung, allogeneic human cord blood derived endothelial cells (HCBEC) were used for the endothelialization of poly-4-methyl-1-pentene (PMP) gas exchange membranes. Therefore, HLA class I expression was silenced stably in HCBECs to prevent rejection. The capacity of HLA class I-silenced HCBEC to abrogate allogeneic immune responses, their functional properties and suitability for endothelialization of PMP membranes were investigated. Delivery of β2-microglobulin (β2m)-specific shRNAs reduced β2m mRNA levels by up to 90% and caused a knockdown of HLA class I expression by up to 85%. HLA-silenced HCBEC abrogated T-cell responses and escaped antibody-mediated complement-dependent cytotoxicity. The EC phenotype and cytokine secretion profiles between HLA-expressing or -silenced HCBEC remained unaltered. EC specific activation (e.g. ICAM) and thrombogenic markers (e.g. thrombomodulin) remained unaffected by HLA-silencing, but their expression was upregulated by TNFα-stimulation. Furthermore, HLA-silenced HCBECs showed high proliferation rates and built an EC monolayer onto PMP membranes. This study represents a new therapeutic concept in the field of cell and organ transplantation and may bring the bioartificial lung as an alternative to lung transplantation closer to reality.

A Simple Method for the Isolation, Cultivation, and Characterization of Endothelial Cells from Rabbit Coronary Circulation

During the last few years, endothelial cell 1.1. Endothelial Cell Harvesting and Culture functions have been intensively studied in several in vitro and in vivo preparations New Zealand white rabbits of either sex (1.5–1.8 [1–3]. However, there are many methodological kg body weight) were anesthetized with a mixture problems that hinder or exclude biochemical analyof xylazine (5 mg/kg) and ketamine (35 mg/kg). ses of intact endothelial cells under in situ condiAnimals received heparin i.v. and 10 minutes later tions. For this reason, a system of cultured coronary were sacrificed with a sodium pentobarbital overendothelial cells grown under physiologically dedose. Then, under careful sterile conditions, the fined and carefully controlled conditions needs to hearts were excised and promptly placed in a sterile be established. Successful cultivation has been rePetri dish. All subsequent work was performed in ported for endothelial cells derived from human a sterile hood under laminar flow conditions. umbilical vein [4], bovine vena cava [5], rabbit pulThe aortas were cannulated with a sterile polymonary artery [6], bovine pulmonary artery [7], ethylene cannula secured with a surgical suture and and from other tissues [8]. Very few studies have the hearts were perfused with 500 mL of sterile described the isolation of endothelial cells from Krebs solution (117 mM NaCl, 6 mM KCl, 3 mM the heart [9,10]; however, the methods described CaCl2, 1 mM MgSO4, 0.5 mM EDTA, 16.7 mM in these reports had limitations represented by a low glucose, 24 mM NaHCO3, pH 7.4) at room temperacellular yield and the need for complex equipment. ture; the Krebs solution was injected manually with Thus, the aim of the present study was to develop a sterile 20-mL syringe to remove all blood compoa method to obtain endothelial cell cultures from nents trapped within the coronary circulation. After the coronary circulation of rabbit hearts, with the this washout, hearts were perfused with 5 mL of a main objectives being a good reproducibility and harvesting solution composed of collagenase type a high cellular yield of pure endothelial cells. IV (Sigma Chemical, St. Louis, MO, USA, cat. no. C-5138, 0.2 U/mL) and trypsin (Sigma Chemical, St. Louis, MO, USA, cat. no. T-8003, 0.3 U/mL) Abbreviations: DMEM, Dulbecco’s modified eagle medium; FCS, dissolved in Krebs solution. After injection of the fetal calf serum; PBS, phosphate-buffered solution; BSA, bovine initial 5 mL, the perfusion was stopped and, beserum albumine; cGMP, guanosine 39:59-cyclic monophospate; ACE, angiotensin converting enzyme. cause the hearts were not beating, the enzyme soluCorresponding author: Paolo Golino, M.D., Ph.D., Division of tion was not washed away, but remained in contact Cardiology, University of Naples “Federico II”, via Sergio Pansini with the coronary vasculature. After 10 minutes of 5, 80131 Naples, Italy. Tel: 139 (81) 746 2216; Fax: 139 (81) 746 2229; E-mail: ,golino@unina.it.. incubation at room temperature, the hearts were

Experimental improvement of microvascular allografts with the new material polyurethane and microvessel endothelial cell seeding

The major aim of this experimental study was the improvement of hemo- and biocompatibility of microvascular allografts. Comparison of polyurethane and ePTFE (expanded polytetrafluoroethene) micrografts did not show a significant advantage of the new material polyurethane. When the endothelial cell seeding method was applied, endothelial cell harvesting from rat fat proved to be very difficult. Therefore, further research on microvessel endothelial cell seeding is planned.

Enzymatic harvesting of adult human saphenous vein endothelial cells: Use of a chemically defined combination of two purified enzymes to attain viable cell yields equal to those attained by crude bacterial collagenase preparations

Seeding vascular prostheses with enzymatically harvested endothelial cells can create endothelial linings that improve small-caliber prosthetic patency. But crude bacterial collagenases used for endothelial harvest contain cytotoxic nonspecific proteases and clostridial cell wall debris which might limit their clinical usefulness. We therefore compared the endothelial cell harvest efficiency of crude bacterial collagenase with that of purified bacterial collagenase alone, purified trypsin alone, and combinations of purified bacterial collagenase and trypsin using concentrations of pure collagenase equal in collagenolytic activity to the crude bacterial collagenase material. The efficiency of harvest from human saphenous vein segments was measured by a microtiter well-growth curve assay of the number of living endothelial cells capable of attachment to fibronectin and subsequent growth obtained per unit area of saphenous vein lumen. Whereas pure collagenase and purified trypsin alone both harvested less than 5% of the baseline endothelial cell density on the veins, a combination of purified collagenase and 0.01% w/v purified trypsin was found to harvest 22% +/- 10% (SD) (n = 8 veins) of the approximately 1.3 x 10(5) endothelial cells/cm2 available on normal saphenous veins. This figure was not statistically different from the harvest efficiency of 19% +/- 10% (N = 4 veins) (p greater than 0.05) obtained by use of 0.1% w/v crude collagenase alone. This result suggests that endothelial harvesting can be done with a defined mixture of pure enzymes which would be clinically preferable to presently used crude extracts of clostridial cultures as a standardized preparation for graft seeding.

Comparison of immediate seeding of endothelial cells with culture lining of small diameter ePTFE carotid interposition grafts

This study is the first to compare chronic healing characteristics of immediately seeded grafts with those of grafts lined by autogenous venous endothelial cells in tissue culture prior to implantation. Ten mongrel dogs had a segment of external jugular vein excised for enzymatic harvest of endothelial cells. After approximately 21 days growth in tissue culture, 4 × 10 6 cells/ml were inoculated into a 6-cm length of 4 mm i.d. ePTFE for formation of a confluent lining in culture media. The remaining external jugular vein had its endothelial cells enzymatically harvested for immediate seeding of an identical length of preclotted ePTFE. Both grafts were implanted end-to-end in the carotid position and excised after 30 days. In 6 of the 10 dogs, grafts were patent bilaterally; all others were occluded. Planimetric measurements on patent grafts with immediate seeding showed a thrombus-free surface area of 56 ± 39% compared to 86 ± 15% for culture-lined grafts ( P = 0.046). Endothelial coverage was 70 ± 24% for immediately seeded grafts and 29 ± 21% for culture-lined grafts ( P = 0.016). We conclude that immediate seeding and culture lining of autogenous endothelial cells in small diameter ePTFE grafts produce equivalent short-term patency. While culture-lined grafts have an initially less thrombogenic luminal surface, subsequent development of a confluent endothelial lining is slower than that with an immediate seeding preparation, and thus would appear to offer no significant clinical benefit, especially in light of the complexity culture lining adds to the procedure.

Effect of Papaverine on Endothelial Cell Harvest from Canine External Jugular Veins

Endothelial cell seeding procedures have been developed to line prosthetic bypass grafts used in peripheral vascular disease; however, because of current inefficient cell harvest techniques a high ratio of vein-to-graft area is necessary. This study was done to determine if the use of papaverine, a smooth muscle cell relaxant, would affect the number or viability of endothelial cells harvested from canine external jugular veins. Using a 0.12 mg/ml solution of papaverine in tissue culture medium to bathe the veins during dissection and excision, the viable cell yield was 2.20 +/- 1.16 (cells x 10(4)/cm2). A control group of veins using standard dissection technique gave a yield of 0.97 +/- 0.40 (p = 0.025). A second group of veins dissected while bathed in tissue culture medium alone gave a yield of 1.82 +/- 0.75, compared to a yield of 2.73 +/- 0.45 for papaverine harvested veins (p = 0.009). Percent cell viability was not significantly different for any of the groups: 73, 70, and 76% for papaverine, control and media only veins, respectively. The papaverine-harvested cells and those harvested with medium alone grew to 95% confluence in tissue culture in 9.8 +/- 1.1 and 9.9 +/- 0.9 days, respectively. Compared to conventional surgical techniques, use of papaverine more than doubled the endothelial cell yield from excised vein segments without adversely affecting viability or rate of growth in cell culture.

Isolation and characterization of endothelial cells from bovine cerebral arteries

For our laboratory's investigation into the role of the endothelial cells in vasospasm following subarachnoid hemorrhage and in inflammatory diseases, we found it necessary to devise a modified method of cell culture, which would be appropriate for studying human endothelial cells from lobectomized brain. We report our techniques to increase cell harvest and ensure reproducibility, our method of culturing endothelial cells from bovine major cerebral arteries, and our morphologic and immunocytochemical characterization of these cells. To increase the harvest of endothelial cells, the blood cells were washed from the lumen of the major cerebral arteries at the slaughterhouse and a modified reversed vessel technique was employed. The monolayer of cultured endothelial cells displayed a cobblestone appearance when it reached confluency and transmission electron microscopy revealed junctional complexes and interdigitation of cytoplasm at Passages 10 and 17. The cells stained positively for Factor VIII-related antigen at Passages 3, 5, 7, 10, and 15. Also the cells metabolized acetylated low-density lipoprotein at Passage 8. To determine the purify of the cultured endothelial cells, an immunocytochemical study of the cytoskeleton was performed on Passage 5 cells using either rhodamine-phalloidin or antibodies against smooth muscle myosin, desmin, and vimentin.

Reduced reproductive capacity of freshly harvested endothelial cells in smokers: A possible shortcoming in the success of seeding?

In an attempt to explain the failure of first clinical trials of autologous endothelial seeding in smokers, the initial reproductive capacity of saphenous vein endothelial cells from smokers and nonsmokers was studied by a replicate microwell technique. Endothelial cells were enzymatically harvested from saphenous vein segments of patients with coronary bypasses (21 smokers and 18 nonsmokers). After 15 minutes (group A) and 7 minutes (group B) of collagenase exposure, the endothelial cell harvest from donors who smoked was 41% (p < 0.02) lower for group A and 30% (p < 0.2) lower for group B than that from nonsmokers. In analogy, the viable cell yield was 32% (p < 0.04) and 29% (p < 0.05) lower for groups A and B, respectively, in cultures from donors who smoked. Daily cell counts over an ensuing 10-day period also revealed a significant difference in the proliferative behavior of endothelial cells from smokers and nonsmokers. Whereas endothelial cells from nonsmokers regularly entered the exponential phase of proliferation on day 4.4 ± 1.8 (group A) and day 4.6 ± 1.3 (group B), endothelial cells from smokers reached the logarithmic growth phase either with delay (day 6.8 ± 2.1, group A) or remained completely quiescent (group B). Lower harvest efficiency and suppressed reproductive capacity of endothelial cells in smokers—on top of an already critically low inoculum in single-staged endothelial cell seeding—might explain the failure of initial clinical trials.

An Improved Procedure for Enzymatic Harvesting of Highly Purified Canine Venous Endothelial Cells for Experimental Small Diameter Vascular Prostheses

We developed a new device, the vein holder, to improve yield and purity of enzymatic harvests of venous endothelial cells. External jugular veins of mongrel dogs were dissected by a no-touch technique. In vitro length and circumference of the vein segments were decreased to about half of the in situ dimensions. The vein holder enabled mounting of the veins at 80% of their in situ length during endothelial cell harvesting. Trypan blue staining and scanning electron microscopic observations revealed that vein eversion as well as the new vein holder technique successfully removed the endothelium. Endothelial cell harvests by the eversion technique were, however, low and varied in size, viability, and purity. In contrast, the defined handling by the new vein holder technique regularly provided markedly increased amounts of endothelial cells. Most of the cells attached and developed cultures consisting of endothelial cells only, as shown by the uptake of DilAcLDL. Prostacyclin production of confluent cultures was similar to that of native veins. It is concluded that minimal handling, defined mounting, and prevention of overfilling the vein markedly improves endothelial cell harvests, providing greater amounts of viable and purified endothelial cells.

The cytotoxic effect of surgical glove powder particles on adult human vascular endothelial cell cultures: Implications for clinical uses of tissue culture techniques

Clinical use of autogenous endothelial cell (EC) seeding of vascular prostheses (VP) would require reliable methods for EC harvest for immediate seeding or primary culture in a hospital or operating room setting. Observation of glove powder particles (GPP) in failed primary adult human saphenous vein EC (AHSVEC) cultures led us to study the effect of surgical GPP on cultured AHSVEC. Addition of GPP to the culture medium of growing ASHVEC cultures reduced the cell counts in a dose-dependent fashion; the mean concentration of GPP required to produce a >50% decrease in cell number was 1.5 ± 0.8 (SD) × 10 4 GPP/ml ( N = 10 experiments), equivalent to a mean dose of 36 μg glove powder per milliliter. The effect was seen within 24 hr of addition of GPP and was not due to interference with EC attachment and spreading or to changes in medium osmolality, pH, glucose, electrolyte, Ca 2+, or Mg 2+ content. Instead, the effect appeared to be due to a filterable toxin added during the final rubber-vulcanizing stage of glove manufacture, since pure cornstarch particles and epichlorhydrin-treated pure cornstarch did not prevent culture growth, whereas 0.2 μm filtrates of medium incubated with GPP taken directly from gloves were lethal. We conclude that filterable cytotoxic substances from GPP may be an avoidable cause of failure in EC seeding of VP, and may affect surgical wound healing as well.

Endothelial cell harvest for seeding vascular prostheses: The influence of technique on cell function, viability, and number

Direct seeding of endothelial cells onto synthetic vascular prostheses has become the subject of increasing surgical research during the last 5 to 7 years. The currently employed cell harvest techniques are inefficient, resulting in cell counts far below the number of cells calculated to be present on the original donor vein. We have compared two methods of enzymatic endothelial cell harvest: cannulation with flushing and eversion over a stainless steel rod. Harvested cells were plated onto tissue culture plastic and counted after 24 hours of incubation. The methods ensured that only those cells viable and functional enough to adhere to the plastic were being considered. Cells were identified as endothelial by immunohistochemical techniques applying antisera to factor VIII-related antigen. Segments of normal vein and of veins treated by each technique were viewed with a scanning electron microscope. Cannulation was the superior method, providing greater numbers of viable, functional cells. The eversion technique was unreliable and probably injurious to endothelial cells.

Endothelial cell harvest for seeding vascular prostheses: The influence of technique on cell function, viability, and number

Endothelial cell harvest for seeding vascular prostheses: The influence of technique on cell function, viability, and number