Comparative Analysis of Blood‐Derived Endothelial Cells for Designing Next‐Generation Personalized Organ‐on‐Chips

Abstract

Background Organ‐on‐chip technology has accelerated in vitro preclinical research of the vascular system, and a key strength of this platform is its promise to impact personalized medicine by providing a primary human cell–culture environment where endothelial cells are directly biopsied from individual tissue or differentiated through stem cell biotechniques. However, these methods are difficult to adopt in laboratories, and often result in impurity and heterogeneity of cells. This limits the power of organ‐chips in making accurate physiological predictions. In this study, we report the use of blood‐derived endothelial cells as alternatives to primary and induced pluripotent stem cell–derived endothelial cells. Methods and Results Here, the genotype, phenotype, and organ‐chip functional characteristics of blood‐derived outgrowth endothelial cells were compared against commercially available and most used primary endothelial cells and induced pluripotent stem cell–derived endothelial cells. The methods include RNA‐sequencing, as well as criterion standard assays of cell marker expression, growth kinetics, migration potential, and vasculogenesis. Finally, thromboinflammatory responses under shear using vessel‐chips engineered with blood‐derived endothelial cells were assessed. Blood‐derived endothelial cells exhibit the criterion standard hallmarks of typical endothelial cells. There are differences in gene expression profiles between different sources of endothelial cells, but blood‐derived cells are relatively closer to primary cells than induced pluripotent stem cell–derived. Furthermore, blood‐derived endothelial cells are much easier to obtain from individuals and yet, they serve as an equally effective cell source for functional studies and organ‐chips compared with primary cells or induced pluripotent stem cell–derived cells. Conclusions Blood‐derived endothelial cells may be used in preclinical research for developing more robust and personalized next‐generation disease models using organ‐on‐chips.

Vascular Transcriptomics: Investigating Endothelial Activation and Vascular Dysfunction Using Blood Outgrowth Endothelial Cells, Organ‐Chips, and RNA Sequencing

“…Our previous studies show that autologous BOECs serve as a suitable alternative to traditionally used primary endothelial cells such as human umbilical vein endothelial cells (HUVECs) or human aortic endothelial cells (HAoECs), as well as patient iPSC-derived endothelial cells (iPSC-ECs) (Mathur et al, 2019;Mathur, Tronolone, & Jain, 2021). In addition to exhibiting disease-specific hallmarks, patient-derived BOECs exhibit patient-specific trends in endothelial activation.…”

“…As an alternative, we have identified an alternative endothelial cell source derived from endothelial progenitors in circulating patient blood (Mathur, Flanagan, & Jain, 2021;Mathur et al, 2019). These blood outgrowth endothelial cells (BOECs) are late-stage endothelial progenitors that are phenotypically similar to other primary endothelial cells and can be isolated relatively easily compared to iPSC-ECs (Mathur, Tronolone, & Jain, 2021). In addition to maintaining a healthy phenotype, BOECs derived from diseased patients are representative of the patient's pathophysiology and can mimic endothelial dysfunction in a patient-specific manner (Mathur, Flanagan, & Jain, 2021).…”

A 3D printable perfused hydrogel vascular model to assay ultrasound-induced permeability

“…However, over the past few years, there has been an increasing interest in utilizing alternative endothelial cell types, such as iPSC-derived endothelial cells and blood-derived endothelial cells as sources that may provide more insight for precision medicine applications. [81][82][83] Future studies will be conducted incorporating these cells with greater therapeutic relevance into this vascular model.…”

Sourcing cells for in vitro models of human vascular barriers of inflammation

“…BOECs are endothelial progenitors isolated from the blood using density gradient isolation and can be matured in culture to create patient-specific EC models ( 69 – 72 ). Their gene expression is similar to HUVECs, and they are responsive to shear stress and cytokine stimulation ( 73 ). Recent studies have incorporated BOECs into tissue chip platforms.…”