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Kolster M, Wilhelmi M, Schrimpf C, et al. Outgrowing endothelial and smooth muscle cells for tissue engineering approaches. J Tissue Eng. 2017;8:2041731417698852doi: 10.1177/2041731417698852.
Kolster, M., Wilhelmi, M., Schrimpf, C., Hilfiker, A., Haverich, A., & Aper, T. (2017). Outgrowing endothelial and smooth muscle cells for tissue engineering approaches. Journal of tissue engineering, 82041731417698852. https://doi.org/10.1177/2041731417698852
Kolster, Moritz, et al. "Outgrowing endothelial and smooth muscle cells for tissue engineering approaches." Journal of tissue engineering vol. 8 (2017): 2041731417698852. doi: https://doi.org/10.1177/2041731417698852
Kolster M, Wilhelmi M, Schrimpf C, Hilfiker A, Haverich A, Aper T. Outgrowing endothelial and smooth muscle cells for tissue engineering approaches. J Tissue Eng. 2017 Mar 15;8:2041731417698852. doi: 10.1177/2041731417698852. eCollection 2017. PMID: 28540031; PMCID: PMC5433677.
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J Tissue Eng. 2017 Mar 15;8:2041731417698852. doi: 10.1177/2041731417698852. eCollection 2017.

Outgrowing endothelial and smooth muscle cells for tissue engineering approaches.

Journal of tissue engineering

Moritz Kolster, Mathias Wilhelmi, Claudia Schrimpf, Andres Hilfiker, Axel Haverich, Thomas Aper

Affiliations

  1. Division of Vascular and Endovascular Surgery, Department of Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School, Hannover, Germany.

PMID: 28540031 PMCID: PMC5433677 DOI: 10.1177/2041731417698852

Abstract

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.

Keywords: Circulating progenitor cells; early outgrowth endothelial cells; endothelial progenitor cells; late outgrowth endothelial cells; late outgrowth smooth muscle cells; tissue engineering

Conflict of interest statement

Declaration of conflicting interests: The authors state that there are commercial associations that might create a conflict of interest including financial interests in connection with this manuscript

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