Development of microfluidics as endothelial progenitor cell capture technology for cardiovascular tissue engineering and diagnostic medicine

Abstract

We have developed a unique microfluidic platform capable of capturing circulating endothelial progenitor cells (EPCs) by understanding surface chemistries and adhesion profiles. The surface of a variable-shear-stress microfluidic device was conjugated with 6 different antibodies [anti-CD34, -CD31, -vascular endothelial growth factor receptor-2 (VEGFR-2), -CD146, -CD45, and -von Willebrand factor (vWF)] designed to match the surface antigens on ovine peripheral blood-derived EPCs. Microfluidic analysis showed a shear-stress-dependent decrease in EPC adhesion on attached surface antigens. EPCs exhibited increased adhesion to antibodies against CD34, VEGFR-2, CD31, and CD146 compared to CD45, consistent with their endothelial cell-specific surface profile, when exposed to a minimum shear stress of 1.47 dyn/cm(2). Bone-marrow-derived mesenchymal stem cells and artery-derived endothelial and smooth muscle cells were used to demonstrate the specificity of the EPC microfluidic device. Coated hematopoietic specific-surface (CD45) and granular vWF antibodies, as well as uncoated bare glass and substrate (1% BSA), were utilized as controls. Microfluidic devices have been developed as an EPC capture platform using immobilized antibodies targeted as EPC surface antigens. This EPC chip may provide a new and effective tool for addressing challenges in cardiovascular disease and tissue engineering.

Clinical Potential of Adult Vascular Progenitor Cells

“…43 Other cardiovascular targets for EPCs include coating of ventricular assist devices and artificial hearts. 44,45 Together, these studies indicate that a number of progenitor cell types exist, with translational potential in therapies as diverse as vasculogenesis augmentation, tissue engineering, hybrid graft/device functionalization, plaque stabilization, modulation of vascular remodeling in pulmonary hypertension, and transplant atherosclerosis prevention. Emerging clinical applications of vascular progenitor cells are illustrated in Figure 2.…”

An Integrated Platform for Isolation, Processing, and Mass Spectrometry-based Proteomic Profiling of Rare Cells in Whole Blood*

“…Previous reports (1)(2)(3)(4), for example, have shown that the concentration of circulating tumor cells (CTCs) within a cancer patient's blood can act as a therapeutic monitoring tool (1)(2)(3)(4). Additionally, the isolation of adult stem cells provides a needed cell source for tissue engineering and regenerative medicine treatments (5,6). Finally, separation and genomic analysis of key cell populations from patients allows for targeted treatment regimens (7,8).…”

Current concepts and new developments for autologous in vivo endothelialisation of biomaterials for intravascular applications

“…After binding, antibody mediated adhesion was not disrupted, even if the shear rate was increased to arterial shear rate. Furthermore, Plouffe et al conjugated the surface of a microfluidic device with antibodies against CD34 or VEGFR-2 to capture in vitro cultivated ovine peripheral blood-derived EPCs (Plouffe et al, 2009). …”