Static spatial growth restriction micropatterning of endothelial colony forming cells influences their morphology and gene expression

Matthew W. Hagen, Monica Hinds

Research output: Contribution to journalArticle

Abstract

Background Endothelialization of small diameter synthetic vascular grafts is a potential solution to the thrombosis and intimal hyperplasia that plague current devices. Endothelial colony forming cells, which are blood-derived and similar to mature endothelial cells, are a potential cell source. Anisotropic spatial growth restriction micropatterning has been previously shown to affect the morphology and function of mature endothelial cells in a manner similar to unidirectional fluid shear stress. To date, endothelial colony forming cells have not been successfully micropatterned. This study addresses the hypothesis that micropatterning of endothelial colony forming cells will induce morphological elongation, cytoskeletal alignment, and changes in immunogenic and thrombogenic–related gene expression. Methods Spatially growth restrictive test surfaces with 25 μm-wide lanes alternating between collagen-I and a blocking polymer were created using microfluidics. Case-matched endothelial colony forming cells and control mature carotid endothelial cells were statically cultured on either micropatterned or non-patterned surfaces. Cell elongation was quantified using shape index. Using confocal microscopy, cytoskeletal alignment was visualized and density and apoptotic rate were determined. Gene expression was measured using quantitative PCR to measure KLF-2, eNOS, VCAM-1, and vWF. Results Endothelial colony forming cells were successfully micropatterned for up to 50 hours. Micropatterned cells displayed elongation and actin alignment. Micropatterning increased the packing densities of both cell types, but did not affect apoptotic rate, which was lower in endothelial colony forming cells. KLF-2 gene expression was increased in micropatterned relative to non-patterned endothelial colony forming cells after 50 hours. No significant differences were seen in the other genes tested. Conclusions Endothelial colony forming cells can be durably micropatterned using spatial growth restriction. Micropatterning has a significant effect on the gross and subcellular morphologies of both cell types. Further study is required to fully understand the effect of micropatterning on endothelial colony forming cell gene expression.

Original languageEnglish (US)
Article numbere0218197
JournalPloS one
Volume14
Issue number6
DOIs
StatePublished - Jun 1 2019

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Gene expression
Endothelial cells
Gene Expression
Elongation
gene expression
Growth
cells
Vascular Cell Adhesion Molecule-1
Confocal microscopy
Microfluidics
Grafts
endothelial cells
Actins
Shear stress
Polymers
Blood
Collagen
Endothelial Cells
Genes
Fluids

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Static spatial growth restriction micropatterning of endothelial colony forming cells influences their morphology and gene expression. / Hagen, Matthew W.; Hinds, Monica.

In: PloS one, Vol. 14, No. 6, e0218197, 01.06.2019.

Research output: Contribution to journalArticle

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