Development of Implantable Autologous Small-Calibre Vascular Grafts from Peripheral Blood Samples

Aper, Thomas; Teebken, Omke E.; Krüger, Alexander; Heisterkamp, Alexander; Hilfiker, Andres; Haverich, Axel

doi:10.1055/s-0032-1315112

Cited by 9 publications

(17 citation statements)

References 20 publications

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“…Complete decellularization, including washing, was performed under agitation at 200 rpm on an orbital shaker. The isolation and cultivation of late outgrowth ovine peripheral blood-derived endothelial cells (oPBEC) was conducted as previously described [Aper et al, 2013]. In brief, 100 mL of peripheral blood from donor sheep (female German black-headed breed, 25-30 kg body weight) were drawn directly from the jugular veins under clean conditions.…”

Section: Decellularization Of Ovvcmentioning

confidence: 99%

Tissue Engineering of Vein Valves Based on Decellularized Natural Matrices

Mogaldea

Goecke²,

Theodoridis³

et al. 2017

Cells Tissues Organs

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Valvular repair or transplantation, designed to restore the venous valve function of the legs, has been proposed as treatment in chronic venous insufficiency. Available grafts or surgeries have provided limited durability so far. Generating venous valve substitutes by means of tissue engineering could be a solution. We generated decellularized jugular ovine vein conduits containing valves (oVVC) after reseeding with ovine endothelial cells differentiated from peripheral blood-derived endothelial cells (oPBEC), cultivated in vitro corresponding to the circulatory situation in the lower leg at rest and under exertion. oVVC were decellularized by detergent treatment. GFP-labeled oPBEC were seeded onto the luminal side of the decellularized oVVC and cultivated under static-rotational conditions for 6 h (group I) and 12 h (group II), respectively. Reseeded matrices of group I were exposed to continuous low flow conditions (“leg at rest”). The tissues of group II were exposed to a gradually increasing flow (“leg under effort”). After 5 days, the grafts of group I revealed a uniform luminal endothelial cell coverage of the examined areas of the venous walls and adjacent venous valve leaflets. In group II, the cell coverage on luminal areas of the venous wall parts was found to be nearly complete. The endothelial cell coverage of adjacent venous valve leaflets was revealed to be less dense and confluent. Endothelial cells cultured on acellular vein tissues of both groups were distinctly orientated uniformly in the flow direction, clearly creating a stable and flow-orientated layer. Thus, an endothelium could successfully be reestablished on the luminal surface of a decellularized venous valve by seeding peripheral blood endothelial cells and culturing under different conditions.

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Section: Decellularization Of Ovvcmentioning

confidence: 99%

Tissue Engineering of Vein Valves Based on Decellularized Natural Matrices

Mogaldea

Goecke²,

Theodoridis³

et al. 2017

Cells Tissues Organs

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“…Den Reifungsprozess in einem Bioreaktor charakterisiert der schrittweise Ersatz der Trägerstruktur durch Matrixproteine, die von den angesiedelten Zellen neu gebildet werden. Insbesondere die Bildung von Kollagen führt zu einer erheblichen Zunahme der Stabilität [6]. Dieser Reifungsprozess ist allerdings aufwendig und zeitintensiv, sodass zusammen mit der notwendigen In-vitro-Vermehrung der Zellen Wochen bis Monate für die Konstruktion eines fertigen Implantats benötigt werden [6].…”

Section: Zelluläre Komponenteunclassified

“…Insbesondere die Bildung von Kollagen führt zu einer erheblichen Zunahme der Stabilität [6]. Dieser Reifungsprozess ist allerdings aufwendig und zeitintensiv, sodass zusammen mit der notwendigen In-vitro-Vermehrung der Zellen Wochen bis Monate für die Konstruktion eines fertigen Implantats benötigt werden [6]. Im Hinblick auf die klinische Nutzung stellt aber die unmittelbare oder zumindest schnelle Verfügbarkeit eine der grundlegenden Anforderungen an ein bioartifizielles Konstrukt dar [23].…”

Section: Zelluläre Komponenteunclassified

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Custom-made autologous bioartificial vascular grafts

Aper

2015

Z Herz- Thorax- Gefäßchir

Self Cite

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“…The most promising are approaches in which cells are seeded on a biological scaffold, which is gradually degraded when seeded cells build a “neotissue.” Nevertheless, although the establishment of coculture settings of EC with SMC has been found to be superior over the seeding of each cell type alone regarding their functionality, at present there is only one description about the use of both EC and SMC isolated and differentiated from peripheral blood probes in TE approaches. 12–15 …”

Section: Introductionmentioning

confidence: 99%

Outgrowing endothelial and smooth muscle cells for tissue engineering approaches

et al. 2017

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In recent years, circulating progenitors of endothelial cells and smooth muscle cells were identified in the peripheral blood. In our study, we evaluated the utilization of both cell types isolated and differentiated from peripheral porcine blood in terms for their use for tissue engineering purposes. By means of density gradient centrifugation, the monocyte fraction from porcine blood was separated, split, and cultivated with specific culture media with either endothelial cell growth medium-2 or smooth muscle cell growth medium-2 for the differentiation of endothelial cells or smooth muscle cells. Obtained cells were characterized at an early stage of cultivation before the first passage and a late stage (fourth passage) on the basis of the expression of the antigens CD31, CD34, CD45, nitric oxide synthase, and the contractile filaments smooth-muscle alpha-actin (sm-alpha-actin) and smoothelin. Functional characterization was done based on the secretion of nitric oxide, the formation of a coherent monolayer on polytetrafluoroethylene, and capillary sprouting. During cultivation in both endothelial cell growth medium-2 and smooth muscle cell growth medium-2, substantially two types of cells grew out: early outgrown CD45-positive cells, which disappeared during further cultivation, and in 85% (n = 17/20) of cultures cultivated with endothelial cell growth medium-2 colony-forming late outgrowth endothelial cells. During cultivation with smooth muscle cell growth medium-2 in 80% (n = 16/20) of isolations colony-forming late outgrowth smooth muscle cells entered the stage. Cultivation with either endothelial cell growth medium-2 or smooth muscle cell growth medium-2 had selective effect on the late outgrown cells to that effect that the number of CD31-positive cells increased from 34.8% ± 13% to 83.9% ± 8% in cultures cultivated with endothelial cell growth medium-2 and the number of sm-α-actin+ cells increased from 52.6% ± 18% to 88% ± 5% in cultures cultivated with smooth muscle cell growth medium-2, respectively. Functional analyses revealed significantly higher levels of nitric oxide secretion, endothelialization capacity, and capillary formation in not expanded cultures cultivated with endothelial cell growth medium-2 in comparison to later stages of cultivation and mature aortic cells. Blood seems to be a reliable and feasible source for the isolation of both endothelial and smooth muscle cells for application in tissue engineering approaches. Whereas, early co-cultures of early and late outgrowth cells provide functional advantages, the differentiation of cells can be directed selectively by the used culture medium for the expansion of highly proliferative late outgrowth endothelial cells and late outgrowth smooth muscle cells, respectively.

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Development of Implantable Autologous Small-Calibre Vascular Grafts from Peripheral Blood Samples

Abstract: Implantable bioartificial vascular grafts can be generated from blood. After cultivation under dynamic conditions the vascular segments possess a structure similar to that of the vascular wall and exhibit biomechanical properties sufficient for implantation as arterial substitutes.

Cited by 9 publications

References 20 publications

Tissue Engineering of Vein Valves Based on Decellularized Natural Matrices

Tissue Engineering of Vein Valves Based on Decellularized Natural Matrices

Custom-made autologous bioartificial vascular grafts

Outgrowing endothelial and smooth muscle cells for tissue engineering approaches

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