Back to Search
Start Over
A pre-conditioning protocol of peripheral blood derived endothelial colony forming cells for endothelialization of tissue engineered constructs
- Source :
- Microvascular Research. 134:104107
- Publication Year :
- 2021
- Publisher :
- Elsevier BV, 2021.
-
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.
- Subjects :
- Male
0301 basic medicine
CD31
Thrombomodulin
Cell Culture Techniques
Pulsatile flow
Neovascularization, Physiologic
CD146 Antigen
030204 cardiovascular system & hematology
Mechanotransduction, Cellular
Biochemistry
Regenerative medicine
Umbilical vein
03 medical and health sciences
Bioreactors
0302 clinical medicine
Antigens, CD
Human Umbilical Vein Endothelial Cells
Humans
Cell Shape
Cells, Cultured
Endothelial Progenitor Cells
Tissue Engineering
Chemistry
Cell Biology
Middle Aged
Cadherins
Actin cytoskeleton
Cell biology
Platelet Endothelial Cell Adhesion Molecule-1
Endothelial stem cell
Glucose
Phenotype
030104 developmental biology
Pulsatile Flow
CD146
Female
Stress, Mechanical
Cardiology and Cardiovascular Medicine
Subjects
Details
- ISSN :
- 00262862
- Volume :
- 134
- Database :
- OpenAIRE
- Journal :
- Microvascular Research
- Accession number :
- edsair.doi.dedup.....aed40d48688243e6b8b281f273ceb8ed
- Full Text :
- https://doi.org/10.1016/j.mvr.2020.104107