Angiogenic potency evaluation of cell therapy candidates by a novel application of the in vitro aortic ring assay

Autor: Peter Szaraz, Farwah Iqbal, Matthew Librach, Andrée Gauthier-Fisher, Clifford Librach
Rok vydání: 2017
Předmět:
0301 basic medicine
Stromal cell
Angiogenesis
Cell- and Tissue-Based Therapy
Mesenchymal stromal cells
Medicine (miscellaneous)
Neovascularization
Physiologic
Aortic ring assay
Biology
In Vitro Techniques
Biochemistry
Genetics and Molecular Biology (miscellaneous)
Cellular regenerative therapy
Umbilical Cord
Cell therapy
lcsh:Biochemistry
Rats
Sprague-Dawley
03 medical and health sciences
Cell Movement
medicine
Animals
Humans
lcsh:QD415-436
Cell migration
Cell Shape
Aorta
Tube formation
lcsh:R5-920
Perivascular cells
Research
Mesenchymal stem cell
Endothelial networks
Endothelial Cells
Mesenchymal Stem Cells
Cell Biology
Coculture Techniques
Cell biology
030104 developmental biology
medicine.anatomical_structure
Microscopy
Fluorescence
Immunology
Molecular Medicine
Biological Assay
Female
Bone marrow
Stem cell
lcsh:Medicine (General)
Pericytes
Zdroj: Stem Cell Research & Therapy
Stem Cell Research & Therapy, Vol 8, Iss 1, Pp 1-14 (2017)
ISSN: 1757-6512
Popis: Background Due to limitations of current angiogenesis assays, we aimed to develop a novel application of the rat aortic ring assay to assess the angiogenic potential of mesenchymal stromal cells (MSCs). First-trimester human umbilical cord-derived perivascular cells (FTM HUCPVCs) have multipotent characteristics and previously demonstrated angiogenic potential. We compared the effect of this young source of MSCs and adult bone marrow stromal cells (BMSCs) on ex vivo aortic endothelial network formation. Methods Thoracic segments of adult rat aortas were isolated, sectioned and embedded into Matrigel™. Fluorophore-labeled FTM HUCPVC lines and BMSCs (N = 3) were cocultured with developing endothelial networks (day 0). MSC integration, tube formation and endothelial network growth were monitored daily using phase-contrast and fluorescence microscopy. Quantification of endothelial networks was performed using ImageJ network analysis software on day 5 of coculture. Results FTM HUCPVCs from two umbilical cord samples migrated toward and integrated with developing aortic ring tubular networks while displaying elongated morphologies (day 1). In contrast, BMSCs did not show targeted migration and maintained spherical morphologies with limited physical interactions. Within 1 week of coculture, FTM HUCPVC lines contributed to significantly greater radial network growth and network loop formation when compared to BMSCs and untreated networks. Conclusions We have developed a novel potency assay to assess the angiogenic potential of cell therapy candidates. Favorable properties of FTM HUCPVCs over BMSCs that we observed with this assay and which merit further study include chemotaxis, affinity for developing vasculature, and physical supportive interactions contributing to the development of endothelial networks. Electronic supplementary material The online version of this article (doi:10.1186/s13287-017-0631-1) contains supplementary material, which is available to authorized users.
Databáze: OpenAIRE
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