A pre-conditioning protocol of peripheral blood derived endothelial colony forming cells for endothelialization of tissue engineered constructs

authored by

Xenia Kraus, Michael Pflaum, Stefanie Thoms, Rebecca Jonczyk, Martin Witt, Thomas Scheper, Cornelia Blume

Abstract

In regenerative medicine, autologous endothelial colony forming cells (ECFCs) bear the greatest potential to be used for surface endothelialization of tissue engineered constructs, as they are easily attainable and possess a high proliferation rate. The aim of this study was to develop a standardized pre-conditioning protocol under dynamic conditions simulating the physiology of human circulation to improve the formation of a flow resistant monolayer of ECFCs and to enhance the antithrombogenicity of the endothelial cells. The main focus of the study was to consequently compare the cellular behavior under a steady laminar flow against a pulsatile flow. Mononuclear cells were isolated out of peripheral blood (PB) buffy coats and plated on uncoated tissue culture flasks in anticipation of guidelines for Advanced Therapy Medicinal Products. ECFCs were identified by typical surface markers such as CD31, CD146 and VE-Cadherin. To explore the effects of dynamic cultivation, ECFCs and human umbilical vein endothelial cells were comparatively cultured under either laminar or pulsatile (1 Hz) flow conditions with different grades of shear stress (5 dyn/cm² versus 20 dyn/cm²). High shear stress of 20 dyn/cm² led to a significant upregulation of the antithrombotic gene marker thrombomodulin in both cell types, but only ECFCs orientated and elongated significantly after shear stress application forming a confluent endothelial cell layer. The work therefore documents a suitable protocol to pre-condition PB-derived ECFCs for sustainable endothelialization of blood contacting surfaces and provides essential knowledge for future cultivations in bioreactor systems.

Organisation(s)

Institute of Technical Chemistry

External Organisation(s)

NIFE - Lower Saxony Centre for Biomedical Engineering, Implant Research and Development
Hannover Medical School (MHH)

Type

Article

Journal

Microvascular research

Volume

134

ISSN

0026-2862

Publication date

03.2021

Publication status

Published

Peer reviewed

Yes

ASJC Scopus subject areas

Biochemistry, Cardiology and Cardiovascular Medicine, Cell Biology

Sustainable Development Goals

SDG 3 - Good Health and Well-being

Electronic version(s)

https://doi.org/10.1016/j.mvr.2020.104107 (Access: Closed)